Compounds for modulating trpv3 function

ABSTRACT

The present application relates to compounds and methods for treating pain and other conditions related to TRPV3.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/752,570, filed Jan. 29, 2013, which is a continuation of U.S. Pat.No. 8,389,546, filed Jul. 1, 2011, which is a continuation of U.S. Pat.No. 7,998,980, filed Sep. 14, 2007, which claims the benefit of priorityto U.S. Provisional Application No. 60/859,139, filed Nov. 15, 2006, andU.S. Provisional Application No. 60/845,039, filed Sep. 15, 2006. Thedisclosures of each of the foregoing applications are herebyincorporated by reference in their entirety.

BACKGROUND

A variety of ion channel proteins exist to mediate ion flux acrosscellular membranes. The proper expression and function of ion channelproteins is essential for the maintenance of cell function,intracellular communication, and the like. Numerous diseases are theresult of misregulation of membrane potential or aberrant calciumhandling. Given the central importance of ion channels in modulatingmembrane potential and ion flux in cells, identification of agents thatcan promote or inhibit particular ion channels are of great interest asresearch tools and as possible therapeutic agents.

One such channel is the Transient Receptor Potential V3 (TRPV3) channel.TRPV3 is a calcium permeable channel, specifically a calcium permeablenon-selective cation channel. In addition to calcium ions, TRPV3channels are permeable to other cations, for example sodium. Thus, TRPV3channels modulate membrane potential by modulating the flux of cationssuch as calcium and sodium ions. Although non-selective cation channelssuch as TRPV3 modulate, among other things, calcium ion flux, they aremechanistically distinct from voltage-gated calcium channels. Generally,voltage-gated calcium channels respond to membrane depolarization andopen to permit an influx of calcium from the extracellular medium thatresults in an increase in intracellular calcium levels orconcentrations. In contrast, TRP channels which are non-selective cationchannels are generally signal transduction gated, long lasting, andproduce more prolonged changes in ion concentration. These mechanisticdifferences are accompanied by structural differences amongvoltage-gated and TRP channels. Thus, although many diverse channels actto regulate ion flux and membrane potential in various cell types and inresponse to numerous stimuli, it is important to recognize thesignificant structural, functional, and mechanistic differences amongdifferent classes of ion channels.

TRPV3 function has been implicated in, among other things, the receptionand transduction of pain. Accordingly, it would be desirable to identifyand make compounds that can modulate one or more functions of TRPV3.Such compounds have a variety of in vitro and in vivo uses.

SUMMARY

An important aspect of achieving cellular homeostasis is the maintenanceof appropriate ion concentrations in various cell types during thedevelopment of and in response to numerous stimuli. Large numbers ofdiverse types of ion channels act to maintain cellular homeostasis bymoving ions into and out of cells across the plasma membrane, and withincells by moving ions across membranes of intracellular organellesincluding, for example, the endoplasmic reticulum, sarcoplasmicreticulum, mitochondria and endocytic organelles including endosomes andlysosomes. One such ion channel is the non-selective cation channelTRPV3. TRPV3 is cation permeable and belongs to the larger family of TRPion channels.

TRP channels have been classified into at least six groups: TRPC(short), TRPV (vanilloid), TRPM (long, melastatin), TRPP (polycystins),TRPML (mucolipins), and TRPA (ANKTM1). The TRPC group can be dividedinto 4 subfamilies (TRPC1, TRPC4,5, TRPC3,6,7 and TRPC2) based onsequence homology and functional similarities. Currently the TRPV familyhas 6 members. TRPV5 and TRPV6 are more closely related to each otherthan to TRPV1, TRPV2, TRPV3, or TRPV4. TRPV3 is most closely related toTRPV4, and is more closely related to TRPV1 and TRPV2 than to TRPV5 andTRPV6. The TRPM family has 8 members. Constituents include thefollowing: the founding member TRPM1 (Melastatin or LTRPC1), TRPM3(KIAA1616 or LTRPC3), TRPM7 (TRP-PLIK, ChaK(1), LTRPC7), TRPM6 (ChaK2),TRPM2 (TRPC7 or LTRPC2), TRPM8 (Trp-p8 or CMR1), TRPM5 (Mtr1 or LTRPC5),and TRPM4 (FLJ20041 or LTRPC4). The sole mammalian member of the TRPAfamily is ANKTM1. The TRPML family consists of the mucolipins, whichinclude TRPML1 (mucolipins 1), TRPML2 (mucolipins 2), and TRPML3(mucolipin3). The TRPP family consists of two groups of channels: thosepredicted to have six transmembrane domains and those that have 11.TRPP2 (PKD2), TRPP3 (PKD2L1), TRPP5 (PKD2L2) are all predicted to havesix transmembrane domains. TRPP1 (PKD1, PC1), PKD-REJ and PKD-1L1 areall thought to have 11 transmembrane domains.

The TRP channels constitute a large and important class of channelsinvolved in modulating cellular homeostasis. The present inventionprovides methods and compositions that modulate at least one TRP familymember. Specifically, the present invention provides methods andcompositions for antagonizing a function of TRPV3. Modulating a functionof TRPV3 provides a means for modulating calcium homeostasis, sodiumhomeostasis, intracellular calcium levels, membrane polarization(resting membrane potential), and/or cation levels in a cell. Compoundsthat can modulate one or more TRPV3 functions are useful in many aspectsincluding, but not limited to, maintaining calcium homeostasis;maintaining sodium homeostasis; modulating intracellular calcium levels;modulating membrane polarization (membrane potential); modulating cationlevels; and/or treating or preventing diseases, disorders, or conditionsassociated with calcium homeostasis, sodium homeostasis, calcium orsodium dyshomeostasis, or membrane polarization/hyperpolarization(including hypo and hyperexcitability), and/or treating or preventingdiseases, disorders, or conditions associated with regulation ormisregulation of TRPV3 expression or function. Additionally, the presentinvention provides, in certain embodiments, methods and compositionsthat antagonize both a function of TRPV3 and a function of one or moreadditional TRP channels.

The present application provides compounds that can modulate TRPV3function. Methods employing these compounds are also provided. Certainembodiments provide a method of modulating a TRPV3 function in a cellcomprising administering to the cell an effective amount of a compoundthat inhibits a TRPV3 mediated current. Certain embodiments provide amethod of modulating a TRPV3 function in a cell comprising administeringto the cell an effective amount of a compound that inhibits TRPV3function, wherein the compound inhibits the Phase II outward currentmediated by TRPV3. Certain embodiments provide a method of preventing ortreating a disease or condition related to TRPV3 function in a subjectcomprising administering to the subject a therapeutically effectiveamount of a compound that inhibits TRPV3 function, wherein the compoundinhibits the Phase II outward current mediated by TRPV3. Certainembodiments provide a method of modulating a TRPV3 function in a cellcomprising administering to the cell an effective amount of a compoundthat inhibits TRPV3 function, wherein the compound inhibits the Phase IIinward current mediated by TRPV3. Certain embodiments also provide amethod of preventing or treating a disease or condition related to TRPV3function in a subject comprising administering to the subject atherapeutically effective amount of a compound that inhibits TRPV3function, wherein the compound inhibits the Phase II inward currentmediated by TRPV3. Certain embodiments provide a method of modulatingTRPV3 function in a cell comprising administering to the cell aneffective amount of a compound that inhibits TRPV3 function, wherein thecompound inhibits the Phase I inward current mediated by TRPV3. Certainembodiments also provide a method of preventing or treating a disease orcondition related to TRPV3 function in a subject comprisingadministering to the subject a therapeutically effective amount of acompound that inhibits TRPV3 function, wherein the compound inhibits thePhase I inward current mediated by TRPV3. Certain embodiments alsoprovide a method of preventing or treating a disease or conditionrelated to TRPV3 function in a subject comprising administering to thesubject a therapeutically effective amount of a compound that inhibitsTRPV3 function, wherein the compound inhibits the Phase I outwardcurrent mediated by TRPV3. Certain embodiments provide a method ofmodulating TRPV3 function in a cell comprising administering to the cellan effective amount of a compound that inhibits TRPV3 function, whereinthe compound inhibits the Phase I outward current mediated by TRPV3.Certain embodiments also provide a method of preventing or treating adisease or condition involving activation of TRPV3 or for which reducedTRPV3 activity can reduce the severity in a subject comprisingadministering to the subject a therapeutically effective amount of acompound that inhibits TRPV3 function, wherein the compound inhibits oneor more of a Phase I inward current mediated by TRPV3, a Phase II inwardcurrent mediated by TRPV3, a Phase I outward current mediated by TRPV3,or a Phase II outward current mediated by TRPV3. In any of theforegoing, the invention additionally provides compounds and methodsthat inhibit both the Phase I outward current and the Phase II outwardcurrent. Furthermore, in any of the foregoing, the invention providescompounds and methods that inhibit both the Phase I inward current andthe Phase II inward current, as well as compounds that inhibit anycombination of Phase I and Phase II currents. Note that inhibition of aparticular current refers to the ability of a compound to inhibit thatcurrent (e.g., Phase I inward, Phase I outward, Phase II inward, and/orPhase II outward) in either an in vitro or in vivo assay Inhibition of aparticular current in either an in vivo or an in vitro assay serves as aproxy for the particular functional activity of the particular compound.

The biphasic currents mediated by TRPV3 are discussed in, for example,Chung et al. (Chung et al., 2005, Journal of Biological Chemistry 280:15928-15941). Briefly, a unique property of TRPV3 is that there is aphase change in the current. The current-voltage relationship changesupon repeated stimulation, so that the amount of inward currentincreases dramatically. For ease, two phases of TRPV3 current have beendescribed: Phase I and Phase II. Throughout, we have defined phase I ascurrents that show a 10:1 ratio or greater of outward current amplitude(at +100 mV) to inward current amplitude (at −120 mV). In other words,the current shows strong outward rectification and minimal inwardcurrent. Phase II is defined as a ration of 2:1 or less of outwardcurrent amplitude (at +100 mV) to inward current amplitude (at −120 mV).The current-voltage relationship is fairly linear in this case.

The following articles are exemplary of the state of the art regardingthe structure and function of TRPV3 (Ramsey et al. (2006) Annual RevPhysiology 68: 619-647; Clapham. (2003) Nature 426: 517-524; Xu et al.(2002) Nature 418: 181-186; Clapham et al. (2001) Nature Reviews ofNeuroscience 2: 387-396). The foregoing articles are incorporated byreference in their entirety.

In one aspect, the invention provides a method of inhibiting a TRPV3function, for example a TRPV3 mediated current or TRPV3 mediated ionflux, in a cell. The method comprises administering to the cell orcontacting the cell with an effective amount of a compound that inhibitsa TRPV3 mediated current with an IC₅₀ less than 10 micromolar. Suitablecompounds include any of the compounds provided herein (e.g., compoundsin Table 1 or compounds have any of the formulae disclosed herein).Suitable TRPV3 antagonist compounds can have any combination of thestructural and/or functional features described herein.

In certain embodiments, the small molecule inhibits a TRPV3 mediatedcurrent or a TRPV3 mediated ipn flux with an IC₅₀ less than 5micromolar, less than 1 micromolar, or less than 500 nanomolar.

In some embodiments, the compounds can be used to inhibit a TRPV3mediated current in vitro, for example in cells in culture. In someembodiments, the compounds can be used to inhibit a TRPV3 mediatedcurrent in vivo. In certain embodiments, the compounds inhibit both aninward and an outward TRPV3-mediated current.

One aspect of the present invention relates to a method for treating orpreventing a condition involving activation of TRPV3 or for whichreduced TRPV3 activity can reduce the severity by administering a TRPV3antagonist that inhibits TRPV3-mediated current. Described in greaterdetail below are TRPV3 antagonists that have measured IC₅₀'s forinhibition of TRPV3 of 10 micromolar or less, 1 micromolar or less, 500nanomolar or less, 200 nanomolar or less, 100 nanomolar or less, andeven 10 nanomolar or less. In certain embodiments, the TRPV3 antagonistinhibit one or both of inward and outward TRPV3-mediated current with anIC₅₀ of 1 micromolar or less, and more preferably with an IC₅₀ of 500nanomolar or less, 200 nanomolar or less, 100 nanomolar or less, 25nanomolar or less and even 10 nanomolar or less. In certain embodiments,the TRPV3 antagonist inhibits at least 95% of TRPV3-mediated current at5 micromolar or less, and even more preferably at 1 micromolar or less.

In certain embodiments, the subject TRPV3 antagonists inhibit TRPV3 withan IC₅₀ at least one order of magnitude lower than its IC₅₀ forinhibition of one or more of TRPV5, TRPV6, NaV 1.2, TRPV1, mitochondrialuniporter and hERG channel activities, and even more preferably at least1.5, two or even three orders of magnitude lower.

In certain embodiments, the subject TRPV3 antagonists inhibit TRPV3 withan IC₅₀ at least one order of magnitude more potent than its Ki for theAMPA receptor. In certain other embodiments, the subject TRPV3antagonists inhibit TRPV3 with an IC₅₀ at least two orders of magnitude,or even three orders of magnitude, or four orders of magnitude morepotent than its Ki for the AMPA receptor. In certain embodiments, thesubject TRPV3 antagonists do not appreciably bind the AMPA receptor. Inother words, the subject antagonists inhibit TRPV3 with a particularIC₅₀ and, when administered at that concentration, the antagonist doesnot appreciably bind AMPA receptor (e.g., does specifically andappreciably bind the AMPA receptor). In certain embodiments, compoundsof the invention inhibit a TRPV3-mediated current with an IC₅₀ that ismore potent than its Ki for the AMPA receptor. In such embodiments, theability of the subject TRPV3 inhibitors to decrease pain would thus beindependent of binding to and modulation of thealpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptorwhich has been implicated in neuropathic pain reception.

In certain embodiments, the TRPV3 antagonists inhibit TRPV3 with an IC₅₀at least one order of magnitude lower than its IC₅₀ for inhibition ofTRPV1, and even more preferably at least 1.5, two or even three ordersof magnitude lower. In certain embodiments, the subject TRPV3antagonists can be selected for selectivity for TRPV3 versus TRPV1 onthe basis of having IC₅₀ for TRPV1 inhibition greater than 10micromolar.

In certain embodiments, the TRPV3 antagonists inhibit one or more ofTRPV2, TRPV4, ANKTM1 and/or TRPM8 with an IC₅₀ of 10 micromolar or less.

In certain embodiments, the TRPV3 antagonist has a therapeutic index(T.I.) for treating the condition with the compound of 10 or greater,and even more preferably has a T.I. of at least 25, 50 or even 100.

In preferred embodiments, the TRPV3 inhibitor has an IC₅₀ for TRPV3inhibition that, at that concentration, does not cause QT intervalelongation in the patient nor alter temperature regulation in thepatient.

In certain embodiments, the TRPV3 inhibitor is used to treat orameliorate pain. Exemplary classes of pain that can treated using aTRPV3 inhibitor include, but are not limited to nociceptive pain,inflammatory pain, and neuropathic pain. Pain that can be treated with aTRPV3 inhibitor can be chronic or acute.

In certain embodiments, the TRPV3 inhibitor is non-narcotic and haslittle or no narcotic side-effects. In certain other embodiments, theTRPV3 inhibitor can be used to treat or ameliorate pain with fewerside-effects than narcotic pain relievers. Exemplary side-effects thatmay be substantially absent at effective dosages of TRPV3 inhibitorsinclude one or more of exopthalmos, catalepsy, disruption of gutmotility, and inhibiton of sensation in non-injured areas of the body.

In certain embodiments, the TRPV3 antagonist is “small molecule”, e.g.,an organic molecule having a molecular weight of 2000 amu or less.Exemplary TRPV3 antagonists include a compound of Formula I or a saltthereof, or a solvate, hydrate, oxidative metabolite or prodrug of thecompound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; mrepresents 0, 1, 2, 3, or 4; R₅ represents a substituent on the ring towhich it is attached; and wherein said compound inhibits TRPV3 with anwith an IC50 of 10 micromolar or less.

In certain embodiments, at least one of R₁ and R₄ represents H.

In certain embodiments, R₁ represents H; R₂, R₃ and R₄ represent H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano; and R₅ is,independently for each occurrence, selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, orcyano.

In certain embodiments, X represents S; R₁ represents H; R₂, R₃ and R₄represent H, lower alkyl, halogen, or CF₃; and m is 0.

In certain embodiments, the TRPV3 antagonist is represented by FormulaIb or a salt thereof, or a solvate, hydrate, oxidative metabolite orprodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; yrepresents 0, 1, 2, or 3; R₅ represents a substituent on the ring towhich it is attached; R₅′ represents a substituent; and wherein saidcompound inhibits TRPV3 with an with an IC50 of 10 micromolar or less.

In certain embodiments, at least one of R₁ and R₄ represents H.

In certain embodiments, R₅′ represents a substituent. In certainembodiments, R₅′ represents lower alkyl or alkoxy.

In certain embodiments, R₁ represents H; R₂, R₃ and R₄ represent H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano; R₅ is, independentlyfor each occurrence, selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano; andR₅′ is selected from lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, X is S; R₁ represents H; R₂, R₃ and R₄ representH, lower alkyl, halogen, cyano, ester, carboxyl, or CF₃; y represents 0;and R₅′ is lower alkyl or alkoxy.

The present invention also relates to certain novel compounds, includingpurified preparations of those compounds. For instance, the inventionprovides compounds of Formula Ib or a salt thereof, or a solvate,hydrate, oxidative metabolite or prodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent wherein at leastone of R₁ and R₄ represents H; X represents S or O; n represents 1 or 2when X is O, or n represents 1 when X is S; y represents 0, 1, 2, or 3;R₅ represents a substituent on the ring to which it is attached; R₅′represents H or a substituent; wherein when R₅′ represents H, R₁represents H; and with the proviso that the compound of Formula Ib isnot the following:

In certain embodiments, the novel compounds of the present invention,including purified preparations of those compounds, are represented byFormula Ib or a salt thereof, or a solvate, hydrate, oxidativemetabolite or prodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent wherein at leastone of R₁ and R₄ represents H; X represents S or O; n represents 1 or 2when X is O, or n represents 1 when X is S; y represents 0, 1, 2, or 3;R₅ represents a substituent on the ring to which it is attached; R₅′represents H or a substituent; wherein when R₅′ represents H, R₁represents H; and with the proviso that when R₅′ is H and y is 0, atleast one of R₁, R₂, R₃, and R₄ is not H, and with the further provisothat when R₅′, R₁, R₂, and R₃ are H and y is 0, R₄ is not heteroaryl orcarboxyl.

In certain embodiments, the novel compounds of the present invention,including purified preparations of those compounds, are represented byFormula Ib or a salt thereof, or a solvate, hydrate, oxidativemetabolite or prodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent wherein at leastone of R₁ and R₄ represents H; X represents S or O; n represents 1 or 2when X is O, or n represents 1 when X is S; y represents 0, 1, 2, or 3;R₅ represents a substituent on the ring to which it is attached; R₅′represents H or a substituent; wherein when R₅′ represents H, R₁represents H; and wherein when R₅′ is H and y is 0, at least one of R₁,R₂, R₃, and R₄ is a substituent, and wherein when y is 0 and R₅′, R₁, R₂and R₃ are H, R₄ represents lower alkyl, alkoxy, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, nitro, halogen, CF₃, aryl, —CH₂CH═CF₂,—CH═CF₂, or cyano.

In certain embodiments, R₅′ represents a substituent. In certainembodiments, R₅′ represents lower alkyl or alkoxy.

In certain embodiments, R₁ represents H; R₂, R₃ and R₄ represent H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano; R₅ is, independentlyfor each occurrence, selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano; andR₅′ is selected from lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, X is S; R₁ represents H; R₂, R₃ and R₄ representH, lower alkyl, halogen, cyano, ester, carboxyl, or CF₃; y represents 0;and R₅′ is lower alkyl or alkoxy.

One aspect of the present invention provides a pharmaceuticalpreparation suitable for use in a human patient, or for veterinary use,comprising an effective amount of any of the compounds shown above(e.g., a compound of Formula I or Ib, or a salt thereof, or a solvate,hydrate, oxidative metabolite or prodrug of the compound or its salt),and one or more pharmaceutically acceptable excipients. In certainembodiments, this pharmaceutical preparation may be for use in treatingor preventing a condition involving activation of TRPV3 or for whichreduced TRPV3 activity can reduce the severity. In certain embodiments,this pharmaceutical preparation may be used for the treatment of adisorder or condition selected from the group consisting of acute orchronic pain, touch sensitivity, burns, inflammation, diabeticneuropathy, psoriasis, eczema, dermatitis, post-herpetic neuralgia(shingles), migraine, incontinence, fever, hot flashes, osteoarthritis,rheumatoid arthritis and cough, or is used as a depilatory to promoteloss of or inhibit the growth of hair on a patient. In certainembodiments, the pharmaceutical preparations have a low enough pyrogenactivity to be suitable for use in a human patient. In certainembodiments, the pharmaceutical preparation comprises an effectiveamount of any of the compounds shown above, wherein the compoundinhibits TRPV3 with an IC₅₀ of 10 micromolar or less. In certainembodiments, the pharmaceutical preparation comprises an effectiveamount of any of the compounds shown above, wherein the compoundinhibits one or more members of the vanilloid receptor family with anIC₅₀ of 10 micromolar or less.

In certain embodiments, the TRPV3 inhibitor for use in methods orpharmaceutical preparations of the present invention is selected from acompound depicted in Table 1. In certain embodiments, the presentinvention contemplates the use of any compound as depicted in Table 1 inany of the methods or pharmaceutical preparations of the presentinvention.

The TRPV3 antagonists of the subject invention can be used as part of aprophylaxis or treatment for a variety of disorders and conditions,including, but not limited to, acute and/or chronic pain, touchsensitivity, burns, inflammation, diabetic neuropathy, psoriasis,eczema, dermatitis, post-herpetic neuralgia (shingles), migraine,incontinence, fever, hot flashes, osteoarthritis, oral mucositis, cancerpain, bladder cystits, pain associated with Crohn's disease andIrritable Bowel Syndrome (IBS), rheumatoid arthritis, Grierson-Gopalansyndrome (better known as burning feet syndrome), burning mouth syndrome(BMS) and cough, or is used as a depilatory to promote loss of orinhibit the growth of hair on a patient. Other exemplary diseases orconditions that can be treated using a TRPV3 antagonist of the presentinvention are detailed throughout the specification. The inventioncontemplates the use of compounds having any of the structures providedin the specification in the treatment of or to reduce the symptoms ofany of the diseases or conditions disclosed in the application. Theinvention further contemplates the use of compounds having any of thestructures provided in the specification in the manufacture of amedicament or pharmaceutical preparation to treat or reduce the symptomsof any of the diseases or conditions provided in the specification.Compounds for use in treating a particular disease or condition can beformulated for administration via a route appropriate for the particulardisease or condition.

The TRPV3 antagonists can be administered alone or in combination withother therapeutic agents. For instance, the TRPV3 antagonist isadministered conjointly with one or more of an anti-inflammatory agent,anti-acne agent, anti-wrinkle agent, anti-scarring agent, anti-psoriaticagent, anti-proliferative agent, anti-fungal agent, anti-viral agent,anti-septic agent, anti-migraine agent, keratolytic agent, or a hairgrowth inhibitor.

The TRPV3 antagonists can be administered topically, orally,transdermally, rectally, vaginally, parentally, intranasally,intraocularly, intravenously, intramuscularly, intraarterially,intrathecally, intracapsularly, intraorbitally, intracardiacly,intradermally, intraperitoneally, transtracheally, subcutaneously,subcuticularly, intraarticularly, subcapsularly, subarachnoidly,intraspinally, intrasternally or by inhallation.

In certain preferred embodiments, the TRPV3 antagonist is administeredtopically.

In certain preferred embodiments, the TRPV3 antagonist is administeredorally.

In certain preferred embodiments, the TRPV3 antagonist is administeredparentally.

In certain preferred embodiments, the TRPV3 antagonist is administeredto prevent, treat or alleviate signs and symptoms of acute pain, chronicpain, touch sensitivity, itching sensitivity, or as part of treating aburn, such as, for example, post-surgical pain, cancer pain, orneuropathic pain.

In certain preferred embodiments, the TRPV3 antagonist is administeredto prevent, treat or alleviate signs and symptoms of migraine.

In certain preferred embodiments, the TRPV3 antagonist is administeredto prevent, treat or alleviate signs and symptoms of a disorder orcondition selected from the group consisting of diabetic neuropathy,inflammation, psoriasis, eczema, dermatitis, post-herpetic neuralgia(shingles), incontinence, bladder incontinence, fever, hot flashes, andcough.

In certain preferred embodiments, the TRPV3 antagonist is administeredto prevent, treat or alleviate signs and symptoms of osteoarthritis.

In certain preferred embodiments, the TRPV3 antagonist is administeredto prevent, treat or alleviate signs and symptoms of rheumatoidarthritis.

In certain preferred embodiments, the TRPV3 antagonist is administeredto prevent, treat or alleviate signs and symptoms of oral mucositis.

In certain preferred embodiments, the TRPV3 antagonist is administeredto promote loss of or inhibit the growth of hair on a patient.

Still another aspect of the present invention relates to the use of aTRPV3 antagonist, e.g., a small molecule agent that inhibits inwardTRPV3-mediated current with an IC₅₀ of 1 micromolar or less, in themanufacture of a medicament to prevent, treat or alleviate symptoms of adisease, disorder or condition involving activation of TRPV3, or forwhich reduced TRPV3 activity can reduce the severity, in a patient.

Yet another aspect of the present invention relates to a pharmaceuticalpreparation comprising an agent that inhibits inward TRPV3-mediatedcurrent with an IC₅₀ of 1 micromolar or less; and a pharmaceuticallyacceptable excipient or solvent wherein the agent is provided in adosage form providing an amount effective to prevent, treat or alleviatesymptoms of a disease, disorder or condition involving activation ofTRPV3, or for which reduced TRPV3 activity can reduce the severity, in apatient. In certain preferred embodiments, the pharmaceuticalpreparation does not cause QT interval elongation in the patient.

In certain illustrative embodiments, the pharmaceutical preparationcomprises an agent that inhibits TRPV3-mediated current with an IC₅₀ ofat least one order of magnitude lower than its IC₅₀ for inhibition ofNaV 1.2 function, TRPV1 function, TRPV5 function, TRPV6 function,mitochondrial uniporter function and HERG function; and apharmaceutically acceptable excipient or solvent, wherein the agent isprovided in a dosage form providing an amount effective to prevent,treat or alleviate symptoms of a disease, disorder or conditioninvolving activation of TRPV3, or for which reduced TRPV3 activity canreduce the severity, in a patient, but which does not cause QT intervalelongation.

In another illustrative embodiment, the pharmaceutical preparationcomprises an agent that inhibits heat-induced TRPV3-mediated currentwith an IC₅₀ of 1 micromolar or less; and a pharmaceutically acceptableexcipient or solvent, wherein the agent is provided in a dosage formproviding an amount effective to prevent, treat or alleviate symptoms ofa disease, disorder or condition involving activation of TRPV3, or forwhich reduced TRPV3 activity can reduce the severity, in a patient, butwhich does not cause QT interval elongation.

One preferred preparation is a topical formulation for reducing TRPV3activity in skin or mucosa, comprising an agent that inhibits both 2-APB(2-aminoethyl diphenylborinate) and heat induced TRPV3-mediated currentwith an IC₅₀ of 1 micromolar or less.

Another preferred preparation is a removable patch or bandage,comprising: (i) a polymeric base; and (ii) an agent that inhibits both2-APB and heat induced TRPV3-mediated current with an IC₅₀ of 1micromolar or less.

Still another illustrative formulation is a skin exfoliant compositionfor topical application to an animal subject comprising a topicalvehicle; one or more skin exfoliant ingredients selected from the groupconsisting of carboxylic acids, keto acids, α-hydroxy acids, β-hydroxyacids, retinoids, peroxides, and organic alcohols, said one or more skinexfoliant ingredients contained in a total amount of at least about 12%by weight and capable of inducing skin irritation and effectingexfoliation of the skin of said subject; and an agent that inhibits both2-APB and heat induced TRPV3-mediated current with an IC₅₀ of 1micromolar or less, which agent is provided in an amount effective foranalgesic, anti-irritant and/or anti-inflammatory effects when appliedto skin.

Yet another embodiment is an antitussive composition for peroraladministration comprising an agent that inhibits both 2-APB and heatinduced TRPV3-mediated current with an IC₅₀ of 1 micromolar or less, andan orally-acceptable pharmaceutical carrier in the form of anaqueous-based liquid, or solid dissolvable in the mouth, selected fromthe group consisting of syrup, elixer, suspension, spray, lozenge,chewable lozenge, powder, and chewable tablet. Such antitussivecompositions can include one or more additional agents for treatingcough, allergy or asthma symptom selected from the group consisting of:antihistamines, 5-lipoxygenase inhibitors, leukotriene inhibitors, H3inhibitors, β-adrenergic receptor agonists, xanthine derivatives,α-adrenergic receptor agonists, mast cell stabilizers, expectorants,NK1, NK2 and NK3 tachykinin receptor antagonists, and GABA_(B) agonists.

Still another embodiment is a metered dose aerosol dispenser containingan aerosol pharmaceutical composition for pulmonary or nasal deliverycomprising an agent that inhibits both 2-APB and heat inducedTRPV3-mediated current with an IC₅₀ of 1 micromolar or less. Forinstance, it can be a metered dose inhaler, a dry powder inhaler or anair-jet nebulizer.

Still another embodiment is an eye ointment or eyedrops for ocularadministration. Such ocular compositions may be useful for the treatmentor alleviation of ocular pain including pain resulting from eye abrasionor post-surgical pain.

In another aspect, the invention contemplates that any of the TRPV3inhibitors of the present invention, including inhibitors having one ormore of the characteristics disclosed herein, can be used to inhibit afunction of TRPV3, for example a TRPV3-mediated current. In someembodiments, the compounds can be used to inhibit a TRPV3 mediatedcurrent in vitro, for example in cells in culture. In some embodiments,the compounds can be used to inhibit a TRPV3 mediated current in vivo.In certain embodiments, the compounds inhibit both an inward and anoutward TRPV3-mediated current.

The invention contemplates pharmaceutical preparations and uses of TRPV3antagonists having any combination of the foregoing or followingcharacteristics, as well as any combination of the structural orfunctional characteristics of the TRPV3 antagonists described herein.Any such antagonists or preparations can be used in the treatment of anyof the diseases or conditions described herein.

DETAILED DESCRIPTION OF THE INVENTION

Cellular homeostasis is a result of the summation of regulatory systemsinvolved in, amongst other things, the regulation of ion flux andmembrane potential. Cellular homeostasis is achieved, at least in part,by movement of ions into and out of cells across the plasma membrane andwithin cells by movement of ions across membranes of intracellularorganelles including, for example, the endoplasmic reticulum,sarcoplasmic reticulum, mitochondria and endocytic organelles includingendosomes and lysosomes.

Movement of ions across cellular membranes is carried out by specializedproteins. TRP channels are one large family of non-selective cationchannels that function to help regulate ion flux and membrane potential.TRP channels are subdivided into 6 sub-families including the TRPV(vanilloid receptor) family. TRPV3 is a member of the TRPV class of TRPchannels.

Non-selective cation channels such as TRPV3 modulate the flux of calciumand sodium ions across cellular membranes. Sodium and calcium influxleads to a depolarization of the cell. This increases the probabilitythat voltage-gated ion channels will reach the threshold required foractivation. As a result, activation of non-selective cation channels canincrease electrical excitability and increase the frequency ofvoltage-dependent events. Voltage dependent events include, but are notlimited to, neuronal action potentials, cardiac action potentials,smooth muscle contraction, cardiac muscle contraction, and skeletalmuscle contraction.

TRPV3 is also highly expressed in skin. In a keratinocyte cell line,stimulation of TRPV3 leads to release of inflammatory mediatorsincluding interleukin-1. Thus TRPV3 may also play an important role inregulating inflammation and pain that results from the release ofinflammatory stimuli (Xu et al., 2006)

TRPV3 proteins are thermosensitive channels expressed in skin cells(see, e.g., Peier et al. (2002) Science 296:2046-2049) and dorsal rootganglion, trigeminal ganglion, spinal cord and brain (see, e.g., Xu etal. (2002) Nature 418:181-185; Smith et al. (2002) Nature 418:186-188).Particular TRPV3 proteins that may be used in screening assays, asdescribed herein, to identify compounds that modulate a function ofTRPV3 include, but are not limited to human TRPV3, mouse TRPV3, andDrosophila TRPV3. U.S. Patent Application Publication 2004/0009537 (the“537 publication”) disclosed sequences corresponding to human, mouse,and Drosophila TRPV3. For example, SEQ ID NOs 106 and 107 of the '537publication correspond to the human nucleic acid and amino acidsequences, respectively. SEQ ID Nos 108 and 109 of the '537 publicationcorrespond to the mouse nucleic acid and amino acid sequences,respectively. The Drosophila protein is approximately 25% identical and41% homologous to the human protein over approximately 49% of the lengthof the protein, and approximately 26% identical and 42% homologous tothe mouse protein over approximately 49% of the length of the protein.

Other exemplary human TRPV3 nucleic acid and amino acid sequences aredisclosed in GenBank at the following accession numbers: gi:21912412(accession no. AJ487035); gi:21435923 (accession no. AF514998);gi:22651775 (accession no. AY118268); gi:85397600 (accession no.BC104868); and gi:22651773 (accession no. AY118267). The TRPV3 sequencesand disclosures provided at these accession numbers are herebyincorporated by reference in their entirety. Compounds that inhibit oneor more functions or activities of TRPV3, according to the presentinvention, inhibit one or more functions of any of the TRPV3 proteinsprovided herein. Furthermore, compounds that inhibit one or morefunctions or activities of TRPV3, according to the present invention,inhibit one or more functions of a TRPV3 protein encoded by a nucleicacid sequence that hybridizes under stringent conditions, including awash step of 0.2×SSC at 65° C., to a TRPV3 nucleic acid sequenceprovided herein.

TRPV3 is expressed in a pattern consistent with a role in, among otherthings, pain. TRPV3 is expressed in tissues containing pain-sensingneurons (nociceptors). Nociceptors mediate responsiveness to force,heat, cold, chemicals, and inflammation. In addition, skin whichexpresses high levels of TRPV3 plays a significant role in pain. Furtherevidence indicates that TRPV3 expression increases in the skin cells ofbreast cancer patients who report significant pain (Gopinath et al.,2005).

Accordingly, modulating the function of TRPV3 proteins provides a meansof modulating calcium homeostasis, sodium homeostasis, membranepolarization, and/or intracellular calcium levels, and compounds thatcan modulate TRPV3 function are useful in many aspects, including, butnot limited to, maintaining calcium homeostasis, modulatingintracellular calcium levels, modulating membrane polarization, andtreating or preventing diseases, disorders, or conditions associatedwith calcium and/or sodium homeostasis or dyshomeostasis. In oneembodiment, compounds that modulate TRPV3 function can be used in thetreatment of diseases, injuries, disorders, or conditions caused orexacerbated, in whole or in part, by regulation or misregulation ofTRPV3 activity. In one embodiment, compounds that inhibit a TRPV3function can be used in the treatment of diseases, injuries, disorders,or conditions caused or exacerbated, in whole or in part, by regulationor misregulation of TRPV3 activity. In still another embodiment,compounds that inhibit a TRPV3 function can be used in the treatment ofpain.

In certain embodiments, the TRPV3 antagonist is “small molecule”, e.g.,an organic molecule having a molecular weight of 2000 amu or less.Exemplary TRPV3 antagonists include a compound of Formula I or a saltthereof, or a solvate, hydrate, oxidative metabolite or prodrug of thecompound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; mrepresents 0, 1, 2, 3, or 4; R₅ represents a substituent on the ring towhich it is attached; and wherein said compound inhibits TRPV3 with anwith an IC50 of 10 micromolar or less.

In certain embodiments, at least one of R₁ and R₄ represents H.

In certain embodiments, R₁ represents H; R₂, R₃ and R₄ represent H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano; and R₅ is,independently for each occurrence, selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, orcyano.

In certain embodiments, X represents S; R₁ represents H; R₂, R₃ and R₄represent H, lower alkyl, halogen, or CF₃; and m is 0.

In certain embodiments, the TRPV3 antagonist is represented by FormulaIb or a salt thereof, or a solvate, hydrate, oxidative metabolite orprodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; yrepresents 0, 1, 2, or 3; R₅ represents a substituent on the ring towhich it is attached; R₅′ represents a substituent; and wherein saidcompound inhibits TRPV3 with an with an IC50 of 10 micromolar or less.

In certain embodiments, at least one of R₁ and R₄ represents H.

In certain embodiments, R₅′ represents a substituent. In certainembodiments, R₅′ represents lower alkyl or alkoxy.

In certain embodiments, R₁ represents H; R₂, R₃ and R₄ represent H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano; R₅ is, independentlyfor each occurrence, selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano; andR₅′ is selected from lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, X is S; R₁ represents H; R₂, R₃ and R₄ representH, lower alkyl, halogen, cyano, ester, carboxyl, or CF₃; y represents 0;and R₅′ is lower alkyl or alkoxy.

Exemplary compounds of Formulae I and Ib are provided in Table 1. Table1 summarizes data collected for the various tested compounds. Table 1provides IC₅₀ data for inhibiting a TRPV3 mediated current. Table 1 alsoprovides selectivity data, where currently available, indicating thedegree to which certain compounds also inhibit other ion channels. Notethat at least two of the compounds represented in Table 1 as inhibitinga TRPV3 mediated current with an IC₅₀ less than or equal to 200 nMinhibit a TRPV3 mediated current with an IC₅₀ of less than or equal to50 nM. Additionally, note that compounds represented in Table 1 havevarious degrees of selectivity for inhibiting a TRPV3 mediated currentin comparison to currents mediated by certain other ion channels.

The present invention also relates to certain novel compounds, includingpurified preparations of those compounds. For instance, the inventionprovides compounds of Formula Ib or a salt thereof, or a solvate,hydrate, oxidative metabolite or prodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent wherein at leastone of R₁ and R₄ represents H; X represents S or O; n represents 1 or 2when X is O, or n represents 1 when X is S; y represents 0, 1, 2, or 3;R₅ represents a substituent on the ring to which it is attached; R₅′represents H or a substituent; wherein when R₅′ represents H, R₁represents H; and with the proviso that the compound of Formula Ib isnot the following:

In certain embodiments, the novel compounds of the present invention,including purified preparations of those compounds, are represented byFormula Ib or a salt thereof, or a solvate, hydrate, oxidativemetabolite or prodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent wherein at leastone of R₁ and R₄ represents H; X represents S or O; n represents 1 or 2when X is O, or n represents 1 when X is S; y represents 0, 1, 2, or 3;R₅ represents a substituent on the ring to which it is attached; R₅′represents H or a substituent; wherein when R₅′ represents H, R₁represents H; and with the proviso that when R₅′ is H and y is 0, atleast one of R₁, R₂, R₃, and R₄ is not H, and with the further provisothat when R₅′, R₁, R₂, and R₃ are H and y is 0, R₄ is not heteroaryl orcarboxyl.

In certain embodiments, the novel compounds of the present invention,including purified preparations of those compounds, are represented byFormula Ib or a salt thereof, or a solvate, hydrate, oxidativemetabolite or prodrug of the compound or its salt:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent wherein at leastone of R₁ and R₄ represents H; X represents S or O; n represents 1 or 2when X is O, or n represents 1 when X is S; y represents 0, 1, 2, or 3;R₅ represents a substituent on the ring to which it is attached; R₅′represents H or a substituent; wherein when R₅′ represents H, R₁represents H; and wherein when R₅′ is H and y is 0, at least one of R₁,R₂, R₃, and R₄ is a substituent, and wherein when y is 0 and R₅′, R₁, R₂and R₃ are H, R₄ represents lower alkyl, alkoxy, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, nitro, halogen, CF₃, aryl, —CH₂CH═CF₂,—CH═CF₂, or cyano.

In certain embodiments, R₅′ represents a substituent. In certainembodiments, R₅′ represents lower alkyl or alkoxy.

In certain embodiments, R₁ represents H; R₂, R₃ and R₄ represent H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano; R₅ is, independentlyfor each occurrence, selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano; andR₅′ is selected from lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, X is S; R₁ represents H; R₂, R₃ and R₄ representH, lower alkyl, halogen, cyano, ester, carboxyl, or CF₃; y represents 0;and R₅′ is lower alkyl or alkoxy.

Exemplary compounds of formula Ib include:

One aspect of the present invention provides a pharmaceuticalpreparation suitable for use in a human patient, or for veterinary use,comprising an effective amount of any of the compounds shown above(e.g., a compound of Formula I or Ib, or a salt thereof, or a solvate,hydrate, oxidative metabolite or prodrug of the compound or its salt),and one or more pharmaceutically acceptable excipients. In certainembodiments, this pharmaceutical preparation may be for use in treatingor preventing a condition involving activation of TRPV3 or for whichreduced TRPV3 activity can reduce the severity. In certain embodiments,this pharmaceutical preparation may be used for the treatment of adisorder or condition selected from the group consisting of acute orchronic pain, touch sensitivity, burns, inflammation, diabeticneuropathy, psoriasis, eczema, dermatitis, post-herpetic neuralgia(shingles), migraine, incontinence, fever, hot flashes, osteoarthritis,rheumatoid arthritis and cough, or is used as a depilatory to promoteloss of or inhibit the growth of hair on a patient. In certainembodiments, the pharmaceutical preparations have a low enough pyrogenactivity to be suitable for use in a human patient. In certainembodiments, the pharmaceutical preparation comprises an effectiveamount of any of the compounds shown above, wherein the compoundinhibits TRPV3 with an IC₅₀ of 10 micromolar or less. In certainembodiments, the pharmaceutical preparation comprises an effectiveamount of any of the compounds shown above, wherein the compoundinhibits one or more members of the vanilloid receptor family with anIC₅₀ of 10 micromolar or less.

In certain embodiments, the TRPV3 inhibitor for use in methods orpharmaceutical preparations of the present invention is depicted inTable 1. The present invention contemplates the use of any compound asdepicted in Table 1. In certain embodiments, the invention contemplatesthat any of the particular compounds depicted in Table 1 can beadministered to treat any of the diseases or conditions disclosedherein. In some embodiments, the compound is formulated as apharmaceutical preparation prior to administration. In certainembodiments, the TRPV3 inhibitor for use in methods or pharmaceuticalpreparations of the present invention is selected from a compounddepicted in Table 1. In certain embodiments, the present inventioncontemplates the use of any compound as depicted in Table 1 in any ofthe methods or pharmaceutical preparations of the present invention.

In certain embodiments of the above formulae, substituents may includeone or more of: alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, orheteroaralkyl, any of which may itself be further substituted, orhalogen, carbonyl (e.g., ester, carboxyl, or formyl), thiocarbonyl(e.g., thioester, thiocarboxylate, or thioformate), ketone, aldehyde, ahydroxyl, an alkoxyl, a sulfhydryl, an alkylthio, amino, acylamino,amido, amidino, cyano, nitro, azido, sulfonyl, sulfoxido, sulfate,sulfonate, sulfamoyl, sulfonamido, a phosphate, and a phosphoryl.

Compounds of any of the above structures may be used in the manufactureof medicaments for the treatment of any diseases disclosed herein.Furthermore, TRPV3 inhibitors, according to the present invention, canbe used in the manufacture of medicaments for the treatment of anydiseases or conditions disclosed herein. In certain embodiments,compounds (e.g., inhibitors) of the invention can be used in thetreatment of pain.

Compounds of any of the above structures may be used to inhibit anactivity of TRPV3 in vitro or in vivo, and/or can be used in themanufacture of medicaments to inhibit an activity of TRPV3 in vitro orin vivo. TRPV3 inhibitors, according to the present invention, can beused to inhibit an activity of TRPV3, and/or can be used in themanufacture of medicaments to inhibit an activity of TRPV3 in vitro orin vivo.

For any of the aspects or embodiments of the invention, an exemplaryfunction of TRPV3 that may be inhibited or modulated by a compound ofthe invention is a TRPV3-mediated current (e.g., an inward or outwardPhase I and/or Phase II current).

In certain aspects, the present invention provides methods for treatingor ameliorating the effects of diseases and conditions using smallmolecules that inhibit a TRPV3-mediated current and/or a TRPV3-mediatedion flux with an IC₅₀ of less than 10 micromolar. Exemplary suitablecompounds for use in any of the methods of the invention (e.g., to treatany of the diseases or conditions disclosed herein) include compoundshaving one or more of the structural or functional characteristicsdisclosed herein (e.g., structure, specificity, potency, solubility,etc.). The present invention contemplates the use of any TRPV3antagonist possessing one or more of the functional or structuralattributes described herein.

In certain embodiments, a suitable compound inhibits an inward and/oroutward TRPV3 mediated current with an IC₅₀ of less than 10 micromolar.In certain embodiments, a suitable compound additionally oralternatively inhibits TRPV3 mediated ion flux with an IC₅₀ of less than10 micromolar. IC₅₀ can be calculated, for example, in an in vitroassay. For example, IC₅₀ can be calculated using electrophysiologicaldeterminations of current, such as standard patch clamp analysis. IC₅₀can also be evaluated using changes in concentration or flux of ionindicators, such as the calcium flux methods described herein.

In particular embodiments, the small molecule is chosen for use becauseit is more selective for one TRP isoform than others, e.g., 10-fold, andmore preferably at least 100- or even 1000-fold more selective for TRPV3over one or more of TRPC6, TRPV5, TRPV6, TRPM8, TRPV1, and/or TRPV4. Inother embodiments, the differential is smaller, e.g., it more stronglyinhibits TRPV3 than TRPM8, TRPV1 and/or TRPV4, preferably at leasttwice, three times, five times, or even ten times more strongly. Suchcomparisons may be made, for example, by comparing IC₅₀ values.

In certain embodiments, a compound which is an antagonist of TRPV3 ischosen to selectively antagonize TRPV3 over other ion channels, e.g.,the compound modulates the activity of TRPV3 at least an order ofmagnitude more strongly than it modulates the activity of NaV1.2,Cav1.2, Cav3.1, HERG, and/or mitochondrial uniporter, preferably atleast two orders of magnitude more strongly, even more preferably atleast three orders of magnitude more strongly. Such comparisons may bemade, for example, by comparing IC₅₀ values.

In certain embodiments, a compound which is an antagonist of TRPV3 ischosen to selectively antagonize TRPV3 over AMPA, e.g., the compoundmodulates the activity of TRPV3 at least an order of magnitude morestrongly than it modulates the activity of AMPA. Such comparisons may bemade, for example, by comparing IC₅₀ values. In certain embodiments, thesubject TRPV3 antagonists do not apperciably bind the AMPA receptor. Inother words, the subject antagonists inhibit TRPV3 with a particularIC₅₀ and, when administered at that concentration, the antagonist doesnot substantially bind the AMPA receptor. In such embodiments, theability of the subject TRPV3 inhibitors to decrease pain would thus beindependent of binding to and modulating thealpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptorwhich has been implicated in neuropathic pain reception. In certainembodiments, the subject TRPV3 antagonists inhibit TRPV3 with an IC₅₀ atleast one order of magnitude more potent than its Ki for the AMPAreceptor. In certain embodiments, the subject TRPV3 antagonists inhibitTRPV3 with an IC₅₀ at least two orders of magnitude more potent than itsKi for the AMPA receptor, or even three or four orders of magnitude morepotent than its Ki for the AMPA receptor.

Similarly, in particular embodiments, the small molecule is chosen foruse because it lacks significant activity against one or more targetsother than TRPV3. For example, the compound may have an IC₅₀ above 500nM, above 1 μM, or even above 10 μM for inhibiting one or more of TRPC6,TRPV5, TRPV6, TRPV1, NaV1.2, Cav1.2, Cav3.1, HERG, and the mitochondrialuniporter.

In certain embodiment, the small molecule is chosen because itantagonizes the function of both TRPV3 and TRPM8, TRPV1 and/or TRPV4.Although such compounds selectively antagonize the function of both ionchannels, the IC₅₀ values need not be identical.

In certain embodiments of any of the foregoing, the small molecule maybe chosen because it is capable of inhibiting heat-induced activation ofTRPV3. In certain embodiments, the TRPV3 antagonist inhibitsheat-induced activation of TRPV3 and 2-APB induced activation of TRPV3.In certain other embodiments, the TRPV3 antagonist inhibits heat-inducedactivation of TRPV3 but does not inhibit 2-APB induced activation ofTRPV3.

In certain embodiments of any of the foregoing, the small molecule maybe chosen because it inhibits a TRPV3 function with an IC₅₀ less than 1uM, or even less than 700, 600, 500, 400, 300, 200, or 100 nM. In otherembodiments, the small molecule is chosen because it inhibits a TRPV3function with an IC₅₀ less than 50 nM, or even less than 25, 20, 10, 5,or 1 nM.

In certain embodiments of any of the foregoing, the compound may bechosen based on the rate of inhibition of a TRPV3 function. In oneembodiment, the compound inhibits a TRPV3 function in less than 5minutes, preferably less than 4, 3, or 2 minutes. In another embodiment,the compound inhibits a TRPV3 function in less than 1 minute.

In any of the foregoing embodiments, the small molecule antagonist ofTRPV3 function may inhibit the Phase I outward current, the Phase Iinward current, the Phase II outward current, the Phase II inwardcurrent, or any combination of one or more of these currents. Compoundsthat inhibit more than one of the foregoing currents may do so with thesame or with differing IC₅₀ values. In any of the foregoing, the abilityof a compound to inhibit a particular Phase I and/or Phase II currentcan be assessed either in vitro or in vivo. Compounds that inhibit anyof the foregoing currents in an in vitro or in vivo assay arecharacterized as compounds that inhibit a function of TRPV3.

In certain embodiments of any of the foregoing, inhibition of a TRPV3function means that a function, for example a TRPV3 mediated current, isdecreased by at least 50% in the presence of an effective amount of acompound in comparison to in the absence of the compound or incomparison to an ineffective amount of a compound. In certain otherembodiments, the inhibition of a TRPV3 function means that a function,for example a TRPV3 mediated current, is decreased by at least 50%, 60%,70%, 75%, 80%, 85%, or 90% in the presence of an effective amount of acompound in comparison to in the absence of the compound. In still otherembodiments, the inhibition of a TRPV3 function means that a function,for example a TRPV3 mediated current, is decreased by at least 92%, 95%,97%, 98%, 98%, 99%, or 100% in the presence of an effective amount of acompound in comparison to in the absence of the compound.

In certain embodiments of any of the foreging, the TRPV3 inhibitor isused to treat or ameliorate pain. Exemplary classes of pain that cantreated using a TRPV3 inhibitor include, but are not limited tonociceptive pain, inflammatory pain, and neuropathic pain. Pain that canbe treated with a TRPV3 inhibitor can be chronic or acute. Throughoutthe specification, a variety of conditions and diseases characterized,at least in part, by pain are discussed in detail. The inventioncontemplates that the pain associated with any of these diseases orconditions can be treated using any of the TRPV3 inhibitors describedherein. The inhibitor can be formulated in a pharmaceutical preparationappropropriate for the intended route of administration.

In certain embodiments of any of the foregoing, the TRPV3 inhibitor canbe used to treat or ameliorate pain with fewer side effects. Forexample, the TRPV3 inhibitor can be used to treat or ameliorate painwithout the narcotic effects of, for example, morphine.

In any of the foregoing or following embodiments, the small molecule canbe characterized by some level of activity versus other ion channels(e.g., certain compounds are selective for inhibiting TRPV3 and othercompounds exhibit a level of cross reactivity against one or more otherion channel). When a small molecule is characterized by its activityagainst another ion channel, inhibition of a function or activity of theother ion channel is defined analogously to the way in which a functionof a TRPV3 channel is defined. Thus, inhibiting the function of anotherion channel means, for example, inhibiting ion flux mediated by thatother ion channel or inhibiting the current mediated by that other ionchannel.

In any of the foregoing embodiments, IC₅₀ values are measured in vitrousing, for example, patch clamp analysis or standard measurements ofcalcium flux. Exemplary in vitro methods that can be used to measureIC₅₀ values of a compound are described in Examples 1 and 2.

Without being bound by theory, a compound may inhibit a function ofTRPV3 by binding covalently or non-covalently to a portion of TRPV3.Alternatively, a compound may inhibit a function of TRPV3 indirectly,for example, by associating with a protein or non-protein cofactornecessary for a function of TRPV3. One of skill in the art will readilyappreciate that an inhibitory compound may associate reversibly orirreversibly with TRPV3 or a cofactor thereof. Compounds that reversiblyassociate with TRPV3 or a cofactor thereof may continue to inhibit afunction of TRPV3 even after dissociation.

The subject TRPV3 inhibitors can be used alone or in combination withother pharmaceutically active agents. Examples of such otherpharmaceutically active agents include, but are not limited to,anti-inflammatory agents (e.g., NSAIDS, hormones and autacoids such ascorticosteroids), anti-acne agents (e.g., retinoids), anti-wrinkleagents, anti-scarring agents, anti-psoriatic agents, anti-proliferativeagents (e.g., anti-eczema agents), anti-fungal agents, anti-viralagents, anti-septic agents (e.g., antibacterials), local anaesthetics,anti-migraine agents, keratolytic agents, hair growth stimulants, hairgrowth inhibitors, and other agents used for the treatment of skindiseases or conditions. Certain active agents belong to more than onecategory.

The subject TRPV3 inhibitors can be used alone or as part of atherapeutic regimen combined with other treatments, therapies, orinterventions appropriate for the particular disease, condition, injuryor disorder being treated. When used as part of a therapeutic regimen,the invention contemplates use of TRPV3 inhibitors in combination withone or more of the following treatment modalities: administration ofnon-TRPV3 inhibitor pharmaceuticals, chemotherapy, radiotherapy,homeopathic therapy, diet, stress management, and surgery.

When administered alone or as part of a therapeutic regimen, in certainembodiments, the invention contemplates administration of TRPV3inhibitors to treat a particular primary disease, injury, disorder, orcondition. Additionally or alternatively, the invention contemplatesadministration of TRPV3 inhibitors to treat pain associated with adisease, injury, disorder, or condition. In still other embodiments, theinvention contemplates administration of TRPV3 inhibitors to treatsymptoms secondary to the primary disease, injury, disorder, orconditions.

The invention contemplates pharmaceutical preparations and uses of TRPV3antagonists having any combination of the foregoing or followingcharacteristics, as well as any combination of the structural orfunctional characteristics of the TRPV3 antagonists described herein.Any such antagonists or preparations can be used in the treatment of anyof the diseases or conditions described herein. Additionally, theinvention contemplates the use of any such antagonists or preparationsfor inhibiting a TRPV3 mediated current in vitro. Combinations of any ofthe foregoing or following aspects and embodiments of the invention arealso contemplated. For example, the invention contemplates that TRPV3antagonists having any of the particular potencies and specificitiesoutlined herein can be formulated for the appropriate route ofadministration and can be used in treating any of the conditions ordiseases detailed herein. In certain embodiments, the inventioncontemplates pharmaceutical preparations and uses of any of the TRPV3antagonists presented in Table 1.

DEFINITIONS

The terms “antagonist” and “inhibitor” are used interchangeably to referto an agent that decreases or suppresses a biological activity, such asto repress an activity of an ion channel, such as TRPV3.

An “effective amount” of, e.g., a TRPV3 antagonist, with respect to thesubject method of treatment, refers to an amount of the antagonist in apreparation which, when applied as part of a desired dosage regimenbrings about a desired clinical or functional result. Without beingbound by theory, an effective amount of a TRPV3 antagonist for use inthe methods of the present invention, includes an amount of a TRPV3antagonist effective to decrease one or more in vitro or in vivofunction of a TRPV3 channel Exemplary functions include, but are notlimited to, intracellular calcium levels, membrane polarization (e.g.,an antagonist may promote hyperpolarization of a cell), Phase I outwardcurrent, Phase II outward current, Phase I inward current, and Phase IIinward current. Compounds that antagonize TRPV3 function includecompounds that antagonize an in vitro or in vivo functional activity ofTRPV3. When a particular functional activity is only readily observablein an in vitro assay, the ability of a compound to inhibit TRPV3function in that in vitro assay serves as a reasonable proxy for theactivity of that compound.

The term “preventing” is art-recognized, and when used in relation to acondition, such as a local recurrence (e.g., pain), a disease such ascancer, a syndrome complex such as heart failure or any other medicalcondition, is well understood in the art, and includes administration ofa composition which reduces the frequency of, or delays the onset of,symptoms of a medical condition in a subject relative to a subject whichdoes not receive the composition. Thus, prevention of cancer includes,for example, reducing the number of detectable cancerous growths in apopulation of patients receiving a prophylactic treatment relative to anuntreated control population, and/or delaying the appearance ofdetectable cancerous growths in a treated population versus an untreatedcontrol population, e.g., by a statistically and/or clinicallysignificant amount. Prevention of an infection includes, for example,reducing the number of diagnoses of the infection in a treatedpopulation versus an untreated control population, and/or delaying theonset of symptoms of the infection in a treated population versus anuntreated control population. Prevention of pain includes, for example,reducing the magnitude of, or alternatively delaying, pain sensationsexperienced by subjects in a treated population versus an untreatedcontrol population.

The present invention provides compounds which are in prodrug form. Theterm “prodrug” is intended to encompass compounds that, underphysiological conditions, are converted into the therapeutically activeagents of the present invention. A common method for making a prodrug isto include selected moieties that are hydrolyzed under physiologicalconditions to reveal the desired molecule. For example, an ester or anamide may be hydrolyzed under physiological conditions to reveal thecorresponding acid. In other embodiments, the prodrug is converted by anenzymatic activity of the host animal. Additionally, prodrugs can beconverted to the compounds of the present invention by chemical orbiochemical methods in an ex vivo environment. For example, prodrugs canbe slowly converted to the compounds of the present invention whenplaced in a transdermal patch reservoir with a suitable enzyme orchemical reagent.

The term “oxidative metabolite” is intended to encompass compounds thatare produced by metabolism of the parent compound under normalphysiological conditions. Specifically, an oxidative metabolite isformed by oxidation of the parent compound during metabolism. Forexample, a thioether group may be oxidized to the correspondingsulfoxide or sulfone.

The term “solvate” as used herein, refers to a compound formed bysolvation (e.g., a compound formed by the combination of solventmolecules with molecules or ions of the solute).

The term “hydrate” as used herein, refers to a compound formed by theunion of water with the parent compound.

The term “treating” includes prophylactic and/or therapeutic treatments.The term “prophylactic or therapeutic” treatment is art-recognized andincludes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic, (i.e., it protects thehost against developing the unwanted condition), whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic, (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The terms “TRPV3”, “TRPV3 protein”, and “TRPV3 channel” are usedinterchangeably throughout the application. These terms refer to an ionchannel (e.g., a polypeptide) comprising the amino acid sequence, forexample, the amino acid sequence of a human TRPV3 protein, or anequivalent polypeptide, or a functional bioactive fragment thereof. Incertain embodiments, the term refers to a polypeptide comprising,consisting of, or consisting essentially of, a TRPV3 amino acid sequenceset forth, for example, in any of the patent applications referencedherein. TRPV3 protein may also include orthologs, for example, mouse,rat, horse, or Drosophila TRPV3.

TRPV3 includes polypeptides that retain a function of TRPV3 and comprise(i) all or a portion of a TRPV3 amino acid sequence (ii) a TRPV3 aminoacid sequence with 1 to about 2, 3, 5, 7, 10, 15, 20, 30, 50, 75 or moreconservative amino acid substitutions; (iii) an amino acid sequence thatis at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%identical to a TRPV3 amino acid sequence; and (iv) functional fragmentsthereof. Polypeptides of the invention also include homologs, e.g.,orthologs and paralogs, of a human TRPV3 polypeptide. TRPV3 polypeptidesand amino acid sequences include, for example, the sequences set forthin any of the patent applications referenced herein.

The term “TRPV3” further refers to a nucleic acid encoding a polypeptideof the invention, e.g., a nucleic acid comprising a sequence consistingof, or consisting essentially of, a TRPV3 polynucleotide sequence. Anucleic acid of the invention may comprise all, or a portion of: (i) aTRPV3 nucleotide sequence; (ii) a nucleotide sequence at least 70%, 75%,80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a TRPV3 nucleotidesequence; (iii) a nucleotide sequence that hybridizes under stringentconditions to a TRPV3 nucleotide sequence; (iv) nucleotide sequencesencoding polypeptides that are functionally equivalent to polypeptidesof the invention; (v) nucleotide sequences encoding polypeptides atleast about 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99% homologous oridentical with a TRPV3 polypeptide sequence; (vi) nucleotide sequencesencoding polypeptides having an activity of a polypeptide of theinvention and having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98%,99% or more homology or identity with a TRPV3 polypeptide sequence;(vii) nucleotide sequences that differ by 1 to about 2, 3, 5, 7, 10, 15,20, 30, 50, 75 or more nucleotide substitutions, additions or deletions,such as allelic variants, of a TRPV3 nucleotide sequence; (viii) nucleicacids derived from and evolutionarily related to a TRPV3 nucleotidesequence; and (ix) complements of, and nucleotide sequences resultingfrom the degeneracy of the genetic code, for all of the foregoing andother nucleic acids of the invention. Nucleic acids of the inventionalso include homologs, e.g., orthologs and paralogs, of a TRPV3 nucleicacid sequence and also variants which have been codon optimized forexpression in a particular organism (e.g., host cell). TRPV3 nucleicacid sequences include, for example, the sequences set forth in any ofthe patent applications referenced herein. Where not explicitly stated,one of skill in the art can readily assess whether TRPV3 refers to anucleic acid or a protein.

The terms “compound” and “agent” are used interchangeably to refer tothe inhibitors/antagonists of the invention. In certain embodiments, thecompounds are small organic or inorganic molecules, e.g., with molecularweights less than 7500 amu, preferably less than 5000 amu, and even morepreferably less than 2000, 1500, 1000, or 500 amu. One class of smallorganic or inorganic molecules are non-peptidyl, e.g., containing 2, 1,or no peptide and/or saccharide linkages. On class of small or organicor inorganic molecules are non-peptidyl and non-nucleic acid containing(e.g., do not include a DNA or RNA moiety). In certain otherembodiments, the compounds are proteins, for example, antibodies oraptamers. Such compounds can bind to and inhibit a function of TRPV3. Incertain other embodiments, the compounds are nucleic acids, for example,TRPV3 antisense oligonucleotides or TRPV3 RNAi constructs. Suchcompounds can inhibit the expression of TRPV3, thereby inhibiting theactivity of TRPV3. Other exemplary compounds that may act as inhibitorsinclude ribozymes and peptide fragments.

The term “acyl” is art-recognized and refers to a group represented bythe general formula hydrocarbylC(O)—, such as alkylC(O)—.

The term “acylamino” is art-recognized and refers to a moiety that canbe represented by the general formula:

wherein R₉ is as defined above, and R′₁₁ represents a hydrogen, analkyl, an alkenyl or —(CH₂)_(m)—R₈, where m and R₈ are as defined above.

The term “acyloxy” is art-recognized and refers to a group representedby the general formula hydrocarbylC(O)O—, such as alkylC(O)O—.

Herein, the term “aliphatic group” refers to a straight-chain,branched-chain, or cyclic aliphatic hydrocarbon group and includessaturated and unsaturated aliphatic groups, such as an alkyl group, analkenyl group, and an alkynyl group.

The terms “alkenyl” and “alkynyl” refer to unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double or triple bond respectively.

The terms “alkoxyl” or “alkoxy” as used herein refers to an alkyl group,as defined below, having an oxygen radical attached thereto.Representative alkoxyl groups include methoxy, ethoxy, propyloxy,tert-butoxy and the like. An “ether” is two hydrocarbons covalentlylinked by an oxygen. Accordingly, the substituent of an alkyl thatrenders that alkyl an ether is or resembles an alkoxyl, such as can berepresented by one of —O-alkyl, —O-alkenyl, —O-alkynyl, —O—(CH₂)_(m)—R₈,where m and R₈ are described above.

The term “alkoxyalkyl” refers to an alkyl group substituted with analkoxy group and may be represented by the general formulaalkyl-O-alkyl.

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups, branched-chain alkyl groups,cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, andcycloalkyl-substituted alkyl groups. In preferred embodiments, astraight chain or branched chain alkyl has 30 or fewer carbon atoms inits backbone (e.g., C₁-C₃₀ for straight chains, C₃-C₃₀ for branchedchains), and more preferably 20 or fewer, and most preferably 10 orfewer. Likewise, preferred cycloalkyls have from 3-10 carbon atoms intheir ring structure, and more preferably have 5, 6 or 7 carbons in thering structure.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification, examples, and claims is intended to include both“unsubstituted alkyls” and “substituted alkyls”, the latter of whichrefers to alkyl moieties having substituents replacing a hydrogen on oneor more carbons of the hydrocarbon backbone. Such substituents caninclude, for example, a halogen, a hydroxyl, a carbonyl (such as acarboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (suchas a thioester, a thioacetate, or a thioformate), an alkoxyl, aphosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, anamido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl,an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, asulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromaticmoiety. It will be understood by those skilled in the art that themoieties substituted on the hydrocarbon chain can themselves besubstituted, if appropriate. For instance, the substituents of asubstituted alkyl may include substituted and unsubstituted forms ofamino, azido, imino, amido, phosphoryl (including phosphonate andphosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl andsulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls(including ketones, aldehydes, carboxylates, and esters), —CF₃, —CN andthe like. Exemplary substituted alkyls are described below. Cycloalkylscan be further substituted with alkyls, alkenyls, alkoxys, alkylthios,aminoalkyls, carbonyl-substituted alkyls, —CF₃, —CN, and the like.

Analogous substitutions can be made to alkenyl and alkynyl groups toproduce, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls,amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls,carbonyl-substituted alkenyls or alkynyls.

Unless the number of carbons is otherwise specified, “lower alkyl” asused herein means an alkyl group, as defined above, but having from oneto ten carbons, more preferably from one to six carbon atoms in itsbackbone structure. Likewise, “lower alkenyl” and “lower alkynyl” havesimilar chain lengths. Throughout the application, preferred alkylgroups are lower alkyls. In preferred embodiments, a substituentdesignated herein as alkyl is a lower alkyl.

The term “C_(x-y)” when used in conjunction with a chemical moiety, suchas, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant toinclude groups that contain from x to y carbons in the chain. C₀alkylindicates a hydrogen where the group is in a terminal position, a bondif internal. A C₁₋₆alkyl group, for example, contains from one to sixcarbon atoms in the chain. The term “alkylamino”, as used herein, refersto an amino group substituted with at least one alkyl group.

The term “alkylthio” refers to an alkyl group, as defined above, havinga sulfur radical attached thereto. In preferred embodiments, the“alkylthio” moiety is represented by one of —S— alkyl, —S-alkenyl,—S-alkynyl, and —S—(CH₂)_(m)—R₈, wherein m and R₈ are defined above.Representative alkylthio groups include methylthio, ethylthio, and thelike.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines, e.g., a moiety that can berepresented by the general formula:

wherein R₉, R₁₀ and R′₁₀ each independently represent a hydrogen, analkyl, an alkenyl, —(CH₂)_(m)—R₈, or R₉ and R₁₀ taken together with theN atom to which they are attached complete a heterocycle having from 4to 8 atoms in the ring structure; R₈ represents an aryl, a cycloalkyl, acycloalkenyl, a heterocycle or a polycycle; and m is zero or an integerin the range of 1 to 8. In preferred embodiments, only one of R₉ or R₁₀can be a carbonyl, e.g., R₉, R₁₀ and the nitrogen together do not forman imide. In certain such embodiments, neither R₉ and R₁₀ is attached toN by a carbonyl, e.g., the amine is not an amide or imide, and the amineis preferably basic, e.g., its conjugate acid has a pK_(a) above 7. Ineven more preferred embodiments, R₉ and R₁₀ (and optionally R′₁₀) eachindependently represent a hydrogen, an alkyl, an alkenyl, or—(CH₂)_(m)—R₈. Thus, the term “alkylamine” as used herein means an aminegroup, as defined above, having a substituted or unsubstituted alkylattached thereto, i.e., at least one of R₉ and R₁₀ is an alkyl group.

The term “aminoalkyl”, as used herein, refers to an alkyl groupsubstituted with an amino group.

The term “amidine” denotes the group —C(NH)—NHR wherein R is H or alkylor aralkyl. In certain embodiments, an amidine is the group —C(NH)—NH₂.

The term “amido” is art-recognized as an amino-substituted carbonyl andincludes a moiety that can be represented by the general formula:

wherein R₉, R₁₀ are as defined above. Preferred embodiments of the amidewill not include imides that may be unstable.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group (e.g., an aromatic or heteroaromatic group).

The term “aryl” as used herein includes 5-, 6-, and 7-memberedsingle-ring aromatic groups that may include from zero to fourheteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazineand pyrimidine, and the like. Those aryl groups having heteroatoms inthe ring structure may also be referred to as “aryl heterocycles”,“heteroaryl” or “heteroaromatics.” In certain embodiments, heteroarylcompounds have one to four heteroatoms in their ring structure, such asone or two heteroatoms. The aromatic ring can be substituted at one ormore ring positions with such substituents as described above, forexample, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphate,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromaticor heteroaromatic moieties, —CF₃, —CN, or the like. The term “aryl” alsoincludes polycyclic ring systems having two or more cyclic rings inwhich two or more carbons are common to two adjoining rings (the ringsare “fused rings”) wherein at least one of the rings is aromatic, e.g.,the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls,aryls and/or heterocyclyls.

The term “carbamate” is art-recognized and refers to a group

wherein R⁹ and R¹⁹ independently represent hydrogen or a hydrocarbylgroup.

The term “carbocycle”, as used herein, refers to an aromatic ornon-aromatic ring in which each atom of the ring is carbon. In certainembodiments, a carbocycle ring contains from 3 to 10 atoms, such as from5 to 7 atoms.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl groupsubstituted with a carbocycle group.

The term “carbonate” is art-recognized and refers to a group —OCO₂—.

The term “carboxy”, as used herein, refers to a group represented by theformula —CO₂H.

The term “carbonyl” is art-recognized and includes such moieties as canbe represented by the general formula:

wherein X is a bond or represents an oxygen or a sulfur, and R₁₁represents a hydrogen, an alkyl, an alkenyl, —(CH₂)_(m)—R₈ or apharmaceutically acceptable salt, R′₁₁ represents a hydrogen, an alkyl,an alkenyl or —(CH₂)_(m)—R₈, where m and R₈ are as defined above. WhereX is an oxygen and R₁₁ or R′₁₁ is not hydrogen, the formula representsan “ester”. Where X is an oxygen, and R₁₁ is as defined above, themoiety is referred to herein as a carboxyl group, and particularly whenR₁₁ is a hydrogen, the formula represents a “carboxylic acid”. Where Xis an oxygen, and R′₁₁ is hydrogen, the formula represents a “formate”.In general, where the oxygen atom of the above formula is replaced bysulfur, the formula represents a “thiocarbonyl” group. Where X is asulfur and R₁₁ or R′₁₁ is not hydrogen, the formula represents a“thioester.” Where X is a sulfur and R₁₁ is hydrogen, the formularepresents a “thiocarboxylic acid.” Where X is a sulfur and R₁₁′ ishydrogen, the formula represents a “thiolformate.” On the other hand,where X is a bond, and R₁₁ is not hydrogen, the above formula representsa “ketone” group. Where X is a bond, and R₁₁ is hydrogen, the aboveformula represents an “aldehyde” group.

The term “electron donating group” refers to chemical groups whichdonate electron density to the atom or group of atoms to which theelectron donating group is attached. The donation of electron densityincludes donation both by inductive and by delocalization/resonanceeffects. Examples of electron donating groups attached to aromatic ringsinclude alkyl, alkenyl, and alkynyl groups, and heteroatoms withelectron lone pairs capable of delocalization.

The term “electron withdrawing group” refers to chemical groups whichwithdraw electron density from the atom or group of atoms to whichelectron withdrawing group is attached. The withdrawal of electrondensity includes withdrawal both by inductive and bydelocalization/resonance effects. Examples of electron withdrawinggroups attached to aromatic rings include perhaloalkyl groups, such astrifluoromethyl, halogens, azides, carbonyl containing groups such asacyl groups, cyano groups, and imine containing groups.

The term “ester”, as used herein, refers to a group —C(O)OR⁹ wherein R⁹represents a hydrocarbyl group.

The term “guanidine” denotes the group —NH—C(NH)—NHR wherein R is H oralkyl or aralkyl. A particular guanidine group is —NH—C(NH)—NH₂.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are boron, nitrogen,oxygen, phosphorus, sulfur and selenium, more preferably nitrogen,oxygen, and sulfur.

The terms “heterocyclyl” or “heterocyclic group” refer to 3- to10-membered ring structures, more preferably 3- to 7-membered rings,whose ring structures include one to four heteroatoms, such as one ortwo heteroatoms. Heterocycles can also be polycycles. Heterocyclylgroups include, for example, thiophene, thianthrene, furan, pyran,isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole,pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine,pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, pyrimidine, phenanthroline, phenazine,phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane,thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactamssuch as azetidinones and pyrrolidinones, sultams, sultones, and thelike. The heterocyclic ring can be substituted at one or more positionswith such substituents as described above, as for example, halogen,alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro,sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl,carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, aheterocyclyl, an aromatic or heteroaromatic moiety, —CF₃, —CN, or thelike.

The term “heterocyclylalkyl”, as used herein, refers to an alkyl groupsubstituted with a heterocycle group.

The term “hydrocarbyl”, as used herein, refers to a group that is bondedthrough a carbon atom that does not have a ═O or ═S substituent, andtypically has at least one carbon-hydrogen bond and a primarily carbonbackbone, but may optionally include heteroatoms. Thus, groups likemethyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are consideredto be hydrocarbyl for the purposes of this application, but substituentssuch as acetyl (which has a ═O substituent on the linking carbon) andethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbylgroups include, but are not limited to aryl, heteroaryl, carbocycle,heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

The term “hydroxyalkyl”, as used herein, refers to an alkyl groupsubstituted with a hydroxy group.

As used herein, the term “nitro” means —NO₂; the term “halogen”designates —F, —Cl, —Br or —I; the term “sulfhydryl” means —SH; the term“hydroxyl” means —OH; and the term “sulfonyl” means —SO₂—.

The terms “polycyclyl” or “polycyclic group” refer to two or more rings(e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/orheterocyclyls) in which two or more carbons are common to two adjoiningrings, e.g., the rings are “fused rings”. In certain embodiments, eachring of the polycycle contains from 3 to 10 atoms in the ring, such asfrom 5 to 7 atoms in the ring. Rings that are joined throughnon-adjacent atoms are termed “bridged” rings. Each of the rings of thepolycycle can be substituted with such substituents as described above,as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic orheteroaromatic moiety, —CF₃, —CN, or the like.

The phrase “protecting group” as used herein means temporarysubstituents which protect a potentially reactive functional group fromundesired chemical transformations. Examples of such protecting groupsinclude esters of carboxylic acids, silyl ethers of alcohols, andacetals and ketals of aldehydes and ketones, respectively. The field ofprotecting group chemistry has been reviewed (Greene, T. W.; Wuts, P. G.M. Protective Groups in Organic Synthesis, 2^(nd) ed.; Wiley: New York,1991).

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and nonaromaticsubstituents of organic compounds (e.g., alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl,heterocyclylalkyl, aralkyl, or heteroaralkyl, any of which may itself befurther substituted), as well as halogen, carbonyl (e.g., ester,carboxyl, or formyl), thiocarbonyl (e.g., thioester, thiocarboxylate, orthioformate), ketone, aldehyde, amino, acylamino, amido, amidino, cyano,nitro, azido, sulfonyl, sulfoxido, sulfate, sulfonate, sulfamoyl,sulfonamido, and phosphoryl. Illustrative substituents include, forexample, those described herein above. The permissible substituents canbe one or more and the same or different for appropriate organiccompounds. For purposes of this invention, the heteroatoms such asnitrogen may have hydrogen substituents and/or any permissiblesubstituents of organic compounds described herein which satisfy thevalences of the heteroatoms. This invention is not intended to belimited in any manner by the permissible substituents of organiccompounds.

It will be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance withpermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc. It will be understood that substitutionon a ring system does not produce a fused polycyclic ring system unlessexplicitely stated.

The term “sulfamoyl” is art-recognized and includes a moiety that can berepresented by the general formula:

in which R₉ and R₁₀ are as defined above.

The term “sulfate” is art recognized and includes a moiety that can berepresented by the general formula:

in which R₄₁ is as defined above.

The term “sulfonamido” is art recognized and includes a moiety that canbe represented by the general formula:

in which R₉ and R′₁₁ are as defined above.

The term “sulfonate” is art-recognized and includes a moiety that can berepresented by the general formula:

in which R₄₁ is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

The terms “sulfoxido” or “sulfinyl”, as used herein, refers to a moietythat can be represented by the general formula:

in which R₄₄ is selected from the group consisting of hydrogen, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl, or aryl.

The term “sulfone” is art-recognized and refers to the group —S(O)₂—.

The term “thioalkyl”, as used herein, refers to an alkyl groupsubstituted with a thiol group.

The term “thioester”, as used herein, refers to a group —C(O)SR⁹ or—SC(O)R⁹ wherein R⁹ represents a hydrocarbyl.

The term “thioether”, as used herein, is equivalent to an ether, whereinthe oxygen is replaced with a sulfur.

The term “urea” is art-recognized and may be represented by the generalformula

wherein R⁹ and R¹⁰ independently represent hydrogen or a hydrocarbyl.

As used herein, the definition of each expression, e.g., alkyl, m, n,etc., when it occurs more than once in any structure, is intended to beindependent of its definition elsewhere in the same structure.

The terms triflyl, tosyl, mesyl, and nonaflyl are art-recognized andrefer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl,and nonafluorobutanesulfonyl groups, respectively. The terms triflate,tosylate, mesylate, and nonaflate are art-recognized and refer totrifluoromethanesulfonate ester, p-toluenesulfonate ester,methanesulfonate ester, and nonafluorobutanesulfonate ester functionalgroups and molecules that contain said groups, respectively.

The abbreviations Me, Et, Ph, Tf, Nf, Ts, Ms represent methyl, ethyl,phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl,p-toluenesulfonyl and methanesulfonyl, respectively. A morecomprehensive list of the abbreviations utilized by organic chemists ofordinary skill in the art appears in the first issue of each volume ofthe Journal of Organic Chemistry; this list is typically presented in atable entitled Standard List of Abbreviations. The abbreviationscontained in said list, and all abbreviations utilized by organicchemists of ordinary skill in the art are hereby incorporated byreference.

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

Methods of preparing substantially isomerically pure compounds are knownin the art. If, for instance, a particular enantiomer of a compound ofthe present invention is desired, it may be prepared by asymmetricsynthesis, or by derivation with a chiral auxiliary, where the resultingdiastereomeric mixture is separated and the auxiliary group cleaved toprovide the pure desired enantiomers. Alternatively, where the moleculecontains a basic functional group, such as amino, or an acidicfunctional group, such as carboxyl, diastereomeric salts may be formedwith an appropriate optically active acid or base, followed byresolution of the diastereomers thus formed by fractionalcrystallization or chromatographic means well known in the art, andsubsequent recovery of the pure enantiomers. Alternatively,enantiomerically enriched mixtures and pure enantiomeric compounds canbe prepared by using synthetic intermediates that are enantiomericallypure in combination with reactions that either leave the stereochemistryat a chiral center unchanged or result in its complete inversion.Techniques for inverting or leaving unchanged a particular stereocenter,and those for resolving mixtures of stereoisomers are well known in theart, and it is well within the ability of one of skill in the art tochoose an appropriate method for a particular situation. See, generally,Furniss et al. (eds.), Vogel's Encyclopedia of Practical OrganicChemistry 5^(th) Ed., Longman Scientific and Technical Ltd., Essex,1991, pp. 809-816; and Heller, Acc. Chem. Res. 23: 128 (1990).

Contemplated equivalents of the compounds described above includecompounds which otherwise correspond thereto, and which have the samegeneral properties thereof (e.g., the ability to inhibit TRPV3activity), wherein one or more simple variations of substituents aremade which do not adversely affect the efficacy of the compound. Ingeneral, the compounds of the present invention may be prepared by themethods illustrated in the general reaction schemes as, for example,described below, or by modifications thereof, using readily availablestarting materials, reagents and conventional synthesis procedures. Inthese reactions, it is also possible to make use of variants which arein themselves known, but are not mentioned here.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Alsofor purposes of this invention, the term “hydrocarbon” is contemplatedto include all permissible compounds having at least one hydrogen andone carbon atom. In a broad aspect, the permissible hydrocarbons includeacyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic organic compounds which can besubstituted or unsubstituted.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

The symbol

, whether utilized as a bond or displayed perpendicular to a bondindicates the point at which the displayed moiety is attached to theremainder of the molecule, solid support, etc.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

Wherein substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents, which would result from writing thestructure from right to left, e.g., —CH₂O— is intended to also recite—OCH₂—; —NHS(O)₂— is also intended to represent —S(O)₂HN—; etc.

The term “pharmaceutically acceptable salts” includes salts of theactive compounds which are prepared with relatively nontoxic acids orbases, depending on the particular substituents found on the compoundsdescribed herein. When compounds of the present invention containrelatively acidic functionalities, base addition salts can be obtainedby contacting the neutral form of such compounds with a sufficientamount of the desired base, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable base addition salts includesodium, potassium, calcium, ammonium, organic amino, or magnesium salt,or a similar salt. When compounds of the present invention containrelatively basic functionalities, acid addition salts can be obtained bycontacting the neutral form of such compounds with a sufficient amountof the desired acid, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable acid addition salts includethose derived from inorganic acids like hydrochloric, hydrobromic,nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, trifluoroacetic, propionic, isobutyric, maleic, malonic,benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic,benzensulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, andthe like. Also included are the salts of amino acids such as arginateand the like, and salts of organic acids like glucuronic or galactunoricacids and the like (see, for example, Berge et al., “PharmaceuticalSalts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certainspecific compounds of the present invention contain both basic andacidic functionalities that allow the compounds to be converted intoeither base or acid addition salts.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers form the various salt forms in certain physical properties, suchas solubility in polar solvents, but otherwise the salts are equivalentto the parent form of the compound for the purposes of the presentinvention.

The term “low enough pyrogen activity”, with reference to apharmaceutical preparation, refers to a preparation that does notcontain a pyrogen in an amount that would lead to an adverse effect(e.g., irritation, fever, inflammation, diarrhea, respiratory distress,endotoxic shock, etc.) in a subject to which the preparation has beenadministered. For example, the term is meant to encompass preparationsthat are free of, or substantially free of, an endotoxin such as, forexample, a lipopolysaccharide (LPS).

Diseases, Disorders, or Conditions Related to TRPV3 Function

In an embodiment of the methods for preventing or treating a disease ordisorder or condition, the agent being administered is one thatmodulates the level and/or activity of a TRPV3 protein. In certainembodiments, the compound inhibits the expression and/or activity of aTRPV3 protein. In other embodiments, the compound selectively inhibitsthe expression of a TRPV3 protein. In other words, in certainembodiment, the compound inhibits the activity of a TRPV3 proteinpreferentially in comparison to the activity of one or more other ionchannels.

In particular embodiments of the methods for preventing or treatingdiseases and disorders provided herein, the disease or disorder can be,for example, a pain or sensitivity to touch such as pain related to adisease or disorder, e.g., cancer pain, a dermatological disease ordisorder, e.g., psoriasis and basal cell and squamous cell cariconomas,a neurodegenerative disease or disorder, e g, Alzheimer's disease (AD),Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis(ALS), and other brain disorders caused by trauma or other insultsincluding aging, an inflammatory disease (e.g., asthma, chronicobstructive pulmonary disease, rheumatoid arthritis, osteoarthritis,inflammatory bowel disease, glomerulonephritis, neuroinflammatorydiseases, multiple sclerosis, and disorders of the immune system),cancer or other proliferative disease, kidney disease and liver disease,a metabolic disorder such as diabetes. Further diseases and conditionsinclude post-surgical pain, post herpetic neuraligia, fibromyalgia, andshingles.

Because of the important role that calcium regulation plays in manycellular processes including cellular activation, gene expression,cellular trafficking and apoptotic cell death, calcium dyshomeostasis isimplicated in the many diseases and disorders involving such cellularactivities. These diseases and disorders include dermatological diseasesand disorders; neurological and neurodegenerative diseases anddisorders; fever associated with various diseases, disorders orconditions; incontinence; inflammatory diseases and disorders such asinflammatory bowel disease and Crohn's disease; respiratory diseases anddisorders such as chronic cough, asthma and chronic obstructivepulmonary disease (COPD); digestive disorders such as ulcers and acidreflux; metabolic diseases and disorders including obesity and diabetes;liver and kidney diseases and disorders; malignancies including cancers;aging-related disorders; and sensitivity to pain and touch.

Additional diseases or conditions that can be treated includeATP-related diseases or disorders including epilepsy, cognition, emesis,pain (e.g., migraine), asthma, peripheral vascular disease,hypertension, immune and inflammatory conditions, irritable bowelsyndrome, cystitis, depression, aging-associated degenerative diseases,urinary incontinence, premature ejaculation, cystic fibrosis, diabetes,contraception and sterility, and wound healing (see, for example,Foresta et al. (1992) J. Biol. Chem. 257:19443-19447; Wang et al. (1990)Biochim Biophys. Res. Commun 166:251-258; Burnstock and Williams, (2000)J. Pharmacol. Exp. Ther. 295: 862-869; and Burnstock, Pharmacol Rev(2006) 58:58-86).

TRPV3 inhibitors described herein can be used in the treatment of any ofthe foregoing or following diseases or conditions, including in thetreatment of pain associated with any of the foregoing or followingdiseases or conditions. When used in a method of treatment, an inhibitorcan be selected and formulated based on the intended route ofadministration.

a. Sensitivity to Pain and Touch, or Pain-Related Diseases or Disorders

Compositions and methods provided herein may be used in connection withprevention or treatment of pain or sensitivity to pain and touch. Painor sensitivity to pain and touch may be indicated in a variety ofdiseases, disorders or conditions, including, but not limited to,diabetic neuropathy, breast pain, psoriasis, eczema, dermatitis, burn,post-herpetic neuralgia (shingles), nociceptive pain, peripheralneuropathic and central neuropathic pain, chronic pain, cancer and tumorpain, spinal cord injury, crush injury and trauma induced pain,migraine, cerebrovascular and vascular pain, Sickle cell disease pain,pancreatitis related pain, rheumatoid arthritis pain, musculoskeletalpain including treating signs and symptoms of osteoarthritis andrheumatoid arthritis, orofacial and facial pain, including dental andcancer related, lower back or pelvic pain, surgical incision relatedpain, inflammatory and non-inflammatory pain, visceral pain, psychogenicpain and soft tissue inflammatory pain, fibromyalgia-related pain, andreflex sympathetic dystrophy (complex regional pain syndrome). Thecompounds and methods of the invention may be used in the treatment ofchronic, as well as acute pain. Chronic or acute pain may be the resultof injury, age, or disease.

Other ion channels have been implicated in reception or transmission ofpain. For example, the involvement of N-type calcium channels in thesynaptic transmissions that convey pain signals from sensory afferentnerve cells to the central nervous system has been recognized. Certainnaturally occurring peptide neurotoxins that specifically block N-typecalcium channel have been shown to act as extremely potent and efficientanalgesics in a wide range of animal pain models, including models ofinflammatory and neuropathic pain. The available evidence suggests thatN-type calcium channel blockers are at least as efficacious as opiates,are devoid of a number of the typical opiate side effects (e.g.respiratory depression) and that the analgesic effect is not subject totolerance development.

It has also been shown that potent peripheral analgesia induced by5-alpha-reduced neurosteroid is mediated in part by effects on T-typeCa²⁺ channels (Pathirathna et al., Pain. 2005 April; 114(3):429-43).

Ethosuximide, an anti-epileptic and relatively selective T-type calciumchannel blocker, has also been shown as being highly effective inreversing neuropathic pain caused by the commonly employed cytotoxicspaclitaxel or vincristine (Flatters and Bennett, Pain. 2004 May;109(1-2):150-61).

Pregabalin, a new drug that interacts with the alpha(2)-delta proteinsubunit of the voltage-gated calcium channel, is an efficacious and safetreatment for the pain of diabetic neuropathy (Richter et al., J Pain.2005 April; 6(4):253-60).

The foregoing demonstrate the involvement of various non-TRP channels inthe reception or transmission of pain. Specifically, the foregoingdemonstrate the involvement of various calcium channels in pain.

TRPV3, as well as TRPV1 and TRPV4, are expressed in a pattern consistentwith involvement in pain. TRPV3 is expressed in pain sensitive neurons,and this expression is upregulated following injury (Smith et al.,2002). In addition, TRPV3 is robustly expressed in skin. Accordingly,methods for treating pain include administration of (i) antagonists of aTRPV3 function; (ii) combinations of selective antagonists of a TRPV3and TRPV1 and/or TRPV4 function; or (iii) a pan-TRP inhibitor thatinhibits a function of TRPV3, TRPV1, and TRPV4.

In addition to TRPV family members, other TRP channels have beenimplicated in pain reception and/or sensation. For example, certain TRPMchannels including TRPM8 have been implicated in the reception and/orsensation of pain. Accordingly, in certain embodiments, the methods ofthe present invention include treating pain by administering (i) acombination of a selective TRPV3 antagonist and a selective TRPM8antagonist; (ii) a combination of a selective TRPV3 antagonist, aselective TRPM8 antagonist, and one or more of a selective TRPV1 and/orTRPV4 antagonist; (iii) a cross-TRP inhibitor that antagonizes afunction of TRPV3 and TRPM8; or (iv) a pan inhibitor that antagonizes afunction of TRPV3, TRPM8, and one or more of TRPV1 and TRPV4.

Without being bound by theory, we propose one possible mechanism for howa TRPV3 antagonist may help reduce pain. TRPV3 antagonists can lead tohyperpolarization of the cell. This may lead to a reduction in thefiring of neurons and/or a decrease in action potential frequency. Inaddition, TRPV3 inhibitors may reduce calcium influx into injured cellsand could prevent the calcium dependent changes in gene expression thatsometimes accompany injury. However, regardless of the mechanism ofaction, available expression analysis, electrophysiology andpharmacological efficacy studies support the use of TRPV3 antagonistsfor the treatment of pain.

These findings are somewhat unanticipated because of the uncertainty andcontroversy generated by analysis of TRPV3 knock out mice. It has beenreported that TRPV3 null mice have deficits in their ability to sensetemperature, but not in their ability to sense pain (Moqrich et al.,2005, Science 307: 1468-1472). This finding contradicted an earlierreport that suggested that TRPV3 null mice had normal thermalthresholds, but were unable to develop thermal hyperalgesia in responseto carrageenan or CFA (Smith et al., 2004, Society for NeuroscienceAbstracts).

b. Dermatological Diseases or Disorders

Influx of calcium across plasma membrane of skin cells is a criticalsignaling element involved in cellular differentiation in the skinepidermis (Dotto, 1999 Crit Rev Oral Biol Med 10:442-457). Regulating ormodulating the calcium entry pathway, and thus a critical control pointfor skin cell growth, can treat or prevent skin diseases or disordersthat are characterized by epidermal hyperplasia, a condition in whichskin cells both proliferate too rapidly and differentiate poorly. Suchdiseases include psoriasis, and basal and squamous cell carcinomas.Psoriasis, estimated to affect up to 7 million Americans, afflictssufferers with mild to extreme discomfort, enhanced susceptibility tosecondary infections, and psychological impact due to disfigurement ofthe affected areas (Lebwohl and Ali, 2001 J Am Acad Dermatol45:487-498). Basal cell carcinomas (BCC) and squamous cell carcinomas(SCC) of the skin represent at least one-third of all cancers diagnosedin the United States each year. More than 1 million new cases arereported annually and incidence is increasing. Despite being relativelynon-aggressive, slow-growing cancers, BCCs are capable of significantlocal tissue destruction and disfigurement. SCCs are more aggressive andthus present even greater complications. Further, given that 80% oflesions are on the head and neck with another 15% on shoulders, back orchest, BCCs and SCCs of the skin can have a significant impact on theappearance and quality of life of the afflicted patient.

Many dermatological disorders are accompanied by itch (pruritus).Pruritus and pain share many mechanistic similarities. Both areassociated with activation of C-fibers, both are potentiated byincreases in temperature and inflammatory mediators and both can bequelled with opiates. Decreasing neuronal excitability, particularlyC-fiber excitability may alleviate pruritus associated with dialysis,dermatitis, pregnancy, poison ivy, allergy, dry skin, chemotherapy andeczema.

Acne is a dermatological disorder of complex etiology. Among otherfactors, secretion of oils from the sebaceous glands that contribute tothe development of acne, Since TRPV3 is also expressed in the sebaceousgland and has been shown to be able to regulate secretion in other skincells, antagonizing TRPV3 function might reduce the signs and symptomsof acne.

c. Neurological or Neurodegenerative Diseases and Disorders

Neurodegenerative diseases and disorders include but are not limited toAlzheimer's disease (AD), Parkinson's disease, Huntington's disease,amyotrophic lateral sclerosis (ALS), and other brain disorders caused bytrauma or other insults including aging.

Mechanisms associated with calcium signaling may be altered in manyneurodegenerative diseases and in disorders resulting from brain injury.For example, fibroblasts or T-lymphocytes from patients with AD haveconsistently displayed an increase in Ca²⁺ release from intracellularstores compared to controls (Ito et al. (1994) Proc. Natl. Acad. Sci.U.S.A. 91:534-538; Gibson et al. (1996) Biochem. Biophys. ACTA1316:71-77; Etchenberrigaray et al. (1998) Neurobiology of Disease,5:37-45). Consistent with these observations, mutations in presenilingenes (PS1 or PS2) associated with familial AD (FAD) have been shown toincrease InsP3-mediated Ca²⁺ release from internal stores (Guo et al.(1996) Neuro Report, 8:379-383; Leissring et al. (1999) J.Neurochemistry, 72:1061-1068; Leissring et al. (1999) J. Biol. Chem.274(46):32535-32538; Leissring et al. (2000) J. Biol. Chem.149(4):793-797; Leissring et al. (2000) Proc. Natl. Acad. Sci. U.S.A.97(15):8590-8593). Furthermore, mutations in PS1 or PS2 associated withan increase in amyloidogenic amyloid β peptide generation in AD arereported to be associated with a decrease in intracellular calcium level(Yoo et al. (2000) Neuron, 27(3):561-572).

Experimental traumatic brain injury has been shown to initiate massivedisturbances in Ca²⁺ concentrations in the brain that may contribute tofurther neuronal damage. Intracellular Ca²⁺ may be elevated by manydifferent ion channels. It has been further shown that channel blockersmay be beneficial in the treatment of neurological motor dysfunctionwhen administered in the acute posttraumatic period (Cheney et al.(2000) J. Neurotrauma, 17(1):83-91).

d. Inflammatory Diseases and Disorders

Compositions and methods provided herein may also be used in connectionwith treatment of inflammatory diseases. These diseases include but arenot limited to asthma, chronic obstructive pulmonary disease, rheumatoidarthritis, osteoarthritis, inflammatory bowel disease,glomerulonephritis, neuroinflammatory diseases such as multiplesclerosis, and disorders of the immune system.

The activation of neutrophils (PMN) by inflammatory mediators is partlyachieved by increasing cytosolic calcium concentration ([Ca²⁺]_(i)).Certain calcium channel-mediated calcium influx in particular is thoughtto play an important role in PMN activation. It has been shown thattrauma increases PMN store-operated calcium influx (Hauser et al. (2000)J. Trauma Injury Infection and Critical Care 48 (4):592-598) and thatprolonged elevations of [Ca²⁺]_(i) due to enhanced store-operatedcalcium influx may alter stimulus-response coupling to chemotaxins andcontribute to PMN dysfunction after injury. Modulation of PMN [Ca²⁺]_(i)through store-operated calcium channels might therefore be useful inregulating PMN-mediated inflammation and spare cardiovascular functionafter injury, shock or sepsis (Hauser et al. (2001) J. Leukocyte Biology69 (1):63-68).

Peripheral neuropathy, for example diabetic neuropathy, is a particularcondition that involves both a neuronal and an inflammatory component.Without being bound by a mechanistic theory, the TRPV3 antagonists ofthe invention may be useful in treating peripheral neuropathiesincluding, but not limited to, diabetic neuropathy. In addition to theiruse in the treatment of peripheral neuropathies (e.g., reducinginflammation), the subject inhibitors may also be useful in reducing thepain associated with peripheral neuropathy.

e. Cancer and Other Proliferative Diseases

Compositions and methods provided herein may also be used in connectionwith treatment of malignancies, including, but not limited to,malignancies of lymphoreticular origin, bladder cancer, breast cancer,colon cancer, endometrial cancer, head and neck cancer, lung cancer,melanoma, ovarian cancer, prostate cancer and rectal cancer, in additionto skin cancers described above. Intracellular calcium level may play animportant role in cell proliferation in cancer cells (Weiss et al.(2001) International Journal of Cancer 92 (6):877-882).

In addition, pain associated with cancer or with cancer treatment is asignificant cause of chronic pain. Cancers of the bone, for example,osteosarcoma, are considered exceptionally painful, and patients withadvanced bone cancer may require sedation to tolerate the intense andpersistent pain. Accordingly, TRPV3 antagonists of the inventionrepresent a significant possible therapeutic for the treatment of pain,for example, the pain associated with cancer or with cancer treatment.

Cancer treatments are not only painful, but they may even be toxic tohealthy tissue. Some chemotherapeutic agents can cause painfulneuropathy. Accordingly, TRPV3 antagonists of the invention represent asignificant possible therapeutic for the treatment of the pain and/orinflammation associated with cancer treatments that cause neuropathy.

A major function of prostaglandins is to protect the gastric mucosa.Included in this function is the modulation of intracellular calciumlevel in human gastric cells which plays a critical role in cellproliferation. Consequently, inhibition of prostaglandins bynonsteroidal anti-inflammatory drugs (NSAIDs) can inhibit calcium influxin gastric cells (Kokoska et al. (1998) Surgery (St Louis) 124(2):429-437). The NSAIDs that relieve inflammation most effectively alsoproduce the greatest gastrointestinal damage (Canadian Family Physician,January 1998, p. 101). Thus, the ability to independently modulatecalcium channels in specific cell types may help to alleviate such sideeffect of anti-inflammatory therapy.

f. Liver Diseases and Disorders

Compositions and methods provided herein may also be used in connectionwith treatment of liver diseases and disorders. These diseases anddisorders include but are not limited to alcoholic liver disease, liverinjury, for example, due to transplantation, hepatitis, cancer, andcirrhosis.

Intracellular calcium level has been implicated in chronic liver disease(Tao et al. (1999) J. Biol Chem., 274(34):23761-23769) as well astransplantation injury after cold preservation-warm reoxygenation(Elimadi et al. (2001) Am J. Physiology, 281(3 Part 1):G809-G815).Chronic ethanol consumption has been shown to impair liver regeneration,in part, by modulating intracellular calcium level (Zhang et al. (1996)J. Clin. Invest. 98(5):1237-1244).

g. Kidney Diseases and Disorders

Compositions and methods provided herein may also be used in connectionwith treatment of kidney diseases and disorders. Mesangial cellhyperplasia is often a key feature of such diseases and disorders. Suchdiseases and disorders may be caused by immunological or othermechanisms of injury, including IgAN, membranoproliferativeglomerulonephritis or lupus nephritis. Imbalances in the control ofmesangial cell replication also appear to play a key role in thepathogenesis of progressive renal failure.

The turnover of mesangial cells in normal adult kidney is very low witha renewal rate of less than 1%. A prominent feature of glomerular/kidneydiseases is mesangial hyperplasia due to elevated proliferation rate orreduced cell loss of mesangial cells. When mesangial cell proliferationis induced without cell loss, for example due to mitogenic stimulation,mesangioproliferative glomerulonephritis can result. Data have indicatedthat regulators of mesangial cell growth, particularly growth factors,may act by regulating certain calcium channels (Ma et al. (2001) J. Am.Soc. of Nephrology, 12:(1) 47-53). Modulators of intracellular calciumlevel may aid in the treatment of glomerular diseases by inhibitingmesangial cell proliferation. The epithelial calcium channel CaT2 hasalso been implicated in hypercalciuria and resultant renal stoneformation (Peng et al. (2000) J. Biol. Chem., 275(36):28186-28194).

h. Incontinence

Incontinence is a significant social and medical problem affecting bothmen and women. Incontinence has many causes including, but not limitedto, age, pregnancy, radiation exposure, surgery, injury, and diseases ofthe bladder or musculature that supports the urethra.

Compositions and methods provided herein may be useful in connectionwith the treatment of incontinence. Animal models of incontinence areoften associated with an increase in the frequency of spontaneous actionpotentials and a chronic depolarization of the smooth muscle cells.Evidence suggests that a non-selective cation current could lead to thisdepolarization. Since TRPV3 mRNA is known to be expressed in bladder, aTRPV3 antagonist may be useful in treating incontinence

i. Temperature Regulation

Because of the effects of ion flux on arterial tension and relaxation,the subject compounds can also be used to affect body temperatureregulation, for example, to reduce fever. Furthermore, given that TRPV3channels are heat responsive channels involved in the reception andsensation of heat stimuli, TRPV3 antagonists can be used to modulate thesensation of heat, warmth, or elevated temperatures.

During menopause, many women experience hot flashes. Hot flashes aremarked by sweating, discomfort, and a generally disproportionateexperience of the temperature of one's environment. The symptoms of hotflashes can be very severe, and may interfere with sleep and other dailyactivities. Furthermore, menopause is not only a condition experiencedby women as they age. Premature menopause, and the accompanyingsymptoms, can be induced by hormonal imbalances, certain medications,cancers or other diseases of the female reproductive tract, and partialor total hysterectomies. Thus, menopause and its symptoms may beexperienced by large numbers of women across a diverse age range.

In certain embodiments, TRPV3 antagonists of the present invention canbe used to decrease the perception of heat and temperature associatedwith hot flashes. TRPV3 antagonists of the present invention can beadministered alone, or as part of a therapeutic regimen to decrease thesymptoms associated with menopause. By way of example, TRPV3 antagonistsof the present invention can be administered alone or together withhormone therapy (e.g., estrogen-replacement therapy) used to decreasethe severity of symptoms associated with menopause.

j. Hypertension

Blockers of voltage-gated calcium channels belong to a class ofmedications originally developed to treat hypertension. Such blockersinhibit the movement of calcium into the muscle cells of the heart andarteries. Because calcium is needed for these muscles to contract, suchblockers lower blood pressure by decreasing the force of cardiaccontractile response and relaxing the muscle walls of the arteries.Although TRPV3 is not a voltage-gated calcium channel, it is stillinstrumental in regulating calcium homeostasis, as well as the balanceof other ions, in cells and tissues. Accordingly, TRPV3 antagonists ofthe invention may be used to treat hypertension. Additional uses of thesubject compounds include other conditions that may be ameliorated, inwhole or in part, by relaxing the muscle walls of blood vessels.Exemplary conditions include headaches and migraine attacks.

k. Hair Loss

TRPV3 knock-out mice have a significant hair loss phenotype.Accordingly, the TRPV3 antagonists of the present invention can be usedto promote hair loss or to otherwise inhibit the growth of body hair Byway of example, TRPV3 antagonists can be applied topically to the skinto promote the loss of or to otherwise inhibit the growth of body hair.In such embodiments, the TRPV3 antagonists act as a depilatory agent topromote the loss of or to otherwise inhibit the growth body hair Whenused in this manner, one or more TRPV3 antagonist can be used alone orin combination with an additional depilatory agent. Additionally, one ormore TRPV3 antagonist can be used to supplement other hair removaltechniques such as waxing or shaving. In such a way, a TRPV3 antagonistcan be used alone or as part of a hair removal regimen to reduce oreliminate unwanted body hair Exemplary unwanted body hair includes, butis not limited to, hair on the legs, arms, back, upper lip, chest,bikini area, underarms, and buttocks.

Additionally or alternatively, TRPV3 antagonists can be administeredsystemically to promote the loss of or to prevent the growth of bodyhair.

In any of the foregoing, TRPV3 antagonists likely provide an improvedmethod for reducing or eliminating unwanted body hair. Given the paininhibiting activity of TRPV3 antagonists, their use alone or as part ofa hair removal regimen provides an improved method for removing bodyhair with less discomfort than currently available waxes and chemicaldepilatories.

As outlined above, compounds that antagonize a function of TRPV3 can beused in the treatment of many diseases, injuries, disorders, andconditions. In certain embodiments, TRPV3 inhibitors can be used in thetreatment of pain. As outlined above, TRPV3 inhibitors can be used inthe treatment of pain resulting from injury or disease, as well as painexperienced as a consequence of treatment. Exemplary classes of paininclude nociceptive pain, inflammatory pain, and neuropathic pain. Suchpain can be chronic or acute. TRPV3 inhibitors can be used in thetreatment of one or more of any of the foregoing classes of pain. Incertain embodiments, TRPV3 inhibitors can be used in the treatment ofnociceptive pain. In certain other embodiments, TRPV3 inhibitors can beused in the treatment of inflammatory pain. In certain otherembodiments, TRPV3 inhibitors can be used in the treatment ofneuropathic pain.

As outlined above, TRPV3 inhibitors may be particularly useful in thetreatment of pain associated with cancer, osteoarthritis, rheumatoidarthritis, post-herpetic neuralgia, burns, and other indicationsdetailed above. To further illustrate, additional exemplary indicationsfor which compounds of the present invention can be used include oralpain, Fabry's disease, complex regional pain syndrome, pancreatitis, andfibromyalgia syndrome.

Fabry's Disease

Vague complaints of pain in hands and feet may be a presenting feature.These symptoms are called acroparesthesias, as they reflect theperipheral neuropathy that is a frequent manifestation of the disease.This pain may be both episodic and chronic. Acute episodes may betriggered by exposure to extremes of temperature, stress, emotion,and/or fatigue.

Fibromyalgia

Fibromyalgia (FMS; fibromyalgia syndrome) is a widespreadmusculoskeletal pain and fatigue disorder. Fibromyalgia is characterizedby pain in the muscles, ligaments, and tendons. The condition affectsmore women than men, and occurs in people of all ages. Overall, FMS isestimated to afflict 3-6% of the population.

Patients have described the pain associated with fibromylagia as deepmuscular aching, throbbing, shooting, and stabbing. The pain sometimesincludes an intense burning sensation. The pain and stiffness are oftenworse in the morning or after repetitive use of a particular musclegroup.

Additionally, varying levels of fatigue ranging from mild toincapacitating are often associated with fibromylagia. Other symptoms offibromylagia include gastrointestinal symptoms. Irritable bowel syndromeand IBS-like symptoms such as constipation, diarrhea, frequent abdominalpain, abdominal gas, and nausea occur in roughly 40 to 70% of FMSpatients. Acid reflux or gastroesophogeal reflux disease (GERD) occursat a similar frequency.

Another frequent and debilitating symptom of FMS is chronic headaches,including migraine and tension-type headaches. Such headaches areexperienced by approximately 70% of FMS patients. Additionally, FMSpatients often experience temporomandibular joint dysfunction syndrome(also known as TMJ) which produces pain in the jaw, teeth, and mouth.TMJ may also exacerbate headaches.

Other common symptoms of FMS include, but are not limited to,premenstrual syndrome and painful periods; chest pain; morningstiffness; cognitive or memory impairment; numbness and tinglingsensations; muscle twitching; irritable bladder; the feeling of swollenextremities; skin sensitivities; dry eyes and mouth; dizziness; andimpaired coordination. Additionally, patients are often sensitive toodors, loud noises, and bright lights.

The cause of FMS remains unknown. However, the onset of the disorder hasbeen linked to infections (viral or bacterial), rheumatoid arthritis,lupus, and hypothyroidism. The link between these and other possibletriggers is unclear.

The impact of FMS on the patient is directly correlated with the levelof pain and fatigue. Pain so severe as to interfere with normal work orfamily functioning. There is currently no cure for FMS, and currenttherapies focus primarily on improving sleep (to decrease fatigue) andtreating pain. Compounds of the present invention could be used to helpmanage the pain associated with FMS. Such pain includes, but is notlimited to, oral pain in the jaw, teeth, and mouth. Such pain alsoincludes non-oral musco-skeletal pain, pain due to headaches, and paindue to gastrointestinal symptoms.

Complex Regional Pain Syndrome (CRPS; also known as chronic regionalpain syndrome) is a chronic pain condition. CRPS was formerly known asreflex sympathetic dystrophy (RSD). CRPS is a chronic, painful, andprogressive neurological condition that affects skin, muscles, joints,and bones. The syndrome usually develops in an injured limb, such as abroken leg or following surgery. However, many cases involve only aminor injury, such as a sprain, and somtimes no precipitating injuriousevent can be identified. CRPS involves continuous, intense pain that isdisproportionate to the severity of the injury. The pain worsens, ratherthan improves, over time.

Although CRPS can affect a variety of regions of the body, it most oftenaffects the arms, legs, hands, or feet. Often the pain begins in oneportion of a limb, but spreads over time to include the entire limb oreven to include a different limb. Typical features include dramaticchanges in the color and temperature of the skin over the affected limbor body part, accompanied by intense burning pain, skin sensitivity,sweating, and swelling.

Generally, CRPS is characterized into two categories. Type I occurs inthe absence of a precipitating nerve injury—although there may have beensome other type of precipitating injury. Type II (formerly calledcausalgia) occurs following a nerve injury. These categories are merelydescriptive, and do not correlate with symptomology or prognosis.

The National Institute of Neurological Disorders and Strokes (NINDS)reports that 2% to 5% of peripheral nerve injury patients and 12% to 21%of patients with paralysis on one side of the body (hemiplegia) developreflex sympathetic dystrophy as a complication. The Reflex SympatheticDystrophy Syndrome Association of America (RSDSA) reports that thecondition occurs following 1-2% of bone fractures.

Precipitating events associated with the onset of CRPS include thefollowing: cerebral lesions, heart disease, heart attack, infection,paralysis on one side of the body (hemiplegia), radiation therapy,repetitive motion disorder (e.g., carpal tunnel syndrome), spinal corddisorders, surgery, and trauma (e.g., bone fracture, gunshot, caraccident). However, in 10-20% of cases, no precipitating event can befound. Note that the injury that precedes the onset of CRPS may or maynot be significant.

The symptoms of CRPS may progress in three stages. An acute stage occursduring the first 1-3 months and may include burning pain, swelling,increased sensitivity to touch, increased hair and nail growth in theaffected region, joint pain, and color and temperature changes. Adystrophic stage may involve constant pain and swelling. The effectedlimb often feels cool to the touch and looks bluish. There is typicallymuscle stiffness and wasting (atrophy), as well as early bone loss(osteoporosis). These symptoms usually occur 3-6 months afterdevelopment of the disorder. During an atrophic stage, the skin becomescool and shiny, increased muscle stiffness and weakness occur, andsymptoms may spread to another limb.

Other symptoms include: burning pain, extreme sensitivity to touch, skincolor changes (red or bluish), skin temperature changes (hot or cold),joint pain, swelling (edema), frequent infections, muscle stiffness,muscle spasm, tremor, weakness, dermatitis, eczema, excessive sweating,and migraine headache. A TRPV3 inhibitor can be useful not only intreating the pain associated with CRPS, but also in relieving many ofthese other symptoms including dermatitis, eczema, and migraines.

Patients with CRPS often suffer from depression and anxiety due to theimpact of the disease of their quality of life.

There is currently no cure for CRPS, and thus treatment typically aimsto relieve painful symptoms. Doctors may prescribe topical analgesics,antidepressants, corticosteroids, and opioids to relieve pain. However,to this point, no single drug or combination of drugs has producedconsistent long-lasting improvement in symptoms. Other treatments mayinclude physical therapy, sympathetic nerve block, spinal cordstimulation, and intrathecal drug pumps to deliver opioids and localanesthetic agents via the spinal cord.

The goals of treatment are to control pain and to maintain as muchmobilization of the affected limb as possible. An individualizedtreatment plan is designed, which often combines treatment modalities.Currently, physical therapy, medications, nerve blocks, and psychosocialsupport are used. TRPV3 inhibitors according to the present inventioncan be used instead of or in addition to one or more of the currenttreatment modalities. For example, a TRPV3 inhibitor can be used as analternative to current medications, but combined with physical therapy.

TRPV3 inhibitors provide an alternative for managing pain in CRPSpatients. TRPV3 inhibitors may be used in combination with any of thecurrent medications used to treat CRPS patients. Alternatively, TRPV3inhibitors may be used as an alternative medication.

In addition to drug therapy, CRPS patients often receive physicaltherapy. TRPV3 inhibitors can be used in addition to physical therapy.Physical therapy may be important for helping retain range of motion andfunction in the affected limb. Appropriate pain management, for exampleusing a TRPV3 inhibitor, not only increases patient comfort, but alsofacilitates involvement in physical therapy.

Regardless of the particular combination of therapies used to managepain in CRPS patients, psychological support is often critical. TRPV3inhibitors can be used in combination with psychological support.

TRPV3 inhibitors of the present invention may be used in the treatmentof CRPS. For example, TRPV3 inhibitors of the present invention may beused to help relieve the pain associated with CRPS. TRPV3 inhibitors canbe used alone or as part of an overall treatment regimen to help managethe pain and other symptoms associated with CRPS. Pain management forCRPS sufferers is critical for maintaining a meaningful quality of life.Furthermore, effective pain management may allow sufferers toparticipate in physical therapy to help retain mobility and use of theeffected limbs.

Pancreatitis is an inflammation of the pancreas. The pancreas is a largegland behind the stomach and close to the duodenum. Normally, digestiveenzymes do not become active until they reach the small intestine, wherethey begin digesting food. But if these enzymes become active inside thepancreas, they start “digesting” the pancreas itself.

Acute pancreatitis occurs suddenly, lasts for a short period of time,and usually resolves. Chronic pancreatitis does not resolve itself andresults in a slow destruction of the pancreas. Either form can causeserious complications including bleeding, tissue damage, and infection.

Acute pancreatitis can be a severe, life-threatening illness with manycomplications. About 80,000 cases occur in the United States each year,and approximately 20 percent of these cases are characterized as severe.

Acute pancreatitis is usually, although not exclusively, caused bygallstones or by alcohol abuse. Acute pancreatitis usually begins withpain in the upper abdomen that may last for a few days. The pain may besevere and may become constant. The pain may be isolated to the abdomenor it may reach to the back and other areas. Sometimes, and for somepatients, the pain is sudden and intense. Other times, or for otherpatients, the pain begins as a mild pain that worsens after eating.Someone with acute pancreatitis often looks and feels very sick. Othersymptoms may include swollen and tender abdomen, nausea, vomiting,fever, and rapid pulse. Severe cases of acute pancreatitis may causedehydration and low blood pressure, and may even lead to organ failure,internal bleeding, or death.

During acute pancreatitis attacks, the blood levels of amylase andlipase are often increased by at least 3-fold. Changes may also occur inblood levels of glucose, calcium, magnesium, sodium, potassium, andbicarbonate.

The current treatment depends on the severity of the attack. Treatment,in general, is designed to support vital bodily functions, manage pain,and prevent complications. Although acute pancreatitis typicallyresolved in a few days, pain management during an attack is oftenrequired. TPRV3 inhibitors can be used to relieve the pain associatedwith acute pancreatitis.

Chronic pancreatitis—If injury to the pancreas continues, chronicpancreatitis may develop. Chronic pancreatitis occurs when digestiveenzymes attack and destroy the pancreas and nearby tissues, causingscarring and pain. Chronic pancreatitis may be caused by alcoholism, orby blocked, damaged, or narrowed pancreatic ducts. Additionally,hereditary factors appear to influence the disease, and in certaincases, there is no identifiable cause (so called idiopathicpancreatitis).

Most people with chronic pancreatitis have abdominal pain. The pain mayget worse when eating or drinking, spread to the back, or becomeconstant and disabling. Other symptoms include nausea, vomiting, weightloss, and fatty stools.

Relieving pain is the first step in treating chronic pancreatitis. Oncethe pain has been managed, a high carbohydrate and low fat dietary planis put in place. Pancreatic enzymes may be used to help compensate fordecrease enzyme production from the injured pancreas. Sometimes insulinor other drugs are needed to control blood glucose.

Although pain is typically managed using drug therapy, surgery may benecessary to relieve pain. Surgery may be necessary to drain an enlargedpancreatic duct or even to removing a portion of a seriously injuredpancreas.

Pain is frequently present with chronic pancreatitis. For example, painis present for approximately 75% of patients with alcoholic chronicpancreatitis, 50% of patients with late-onset idiopathic chronicpancreatitis, and 100% of patients with early-onset idiopathic chronicpancreatitis (DiMagno, 1999, Gastroenterology 116(5): 1252-1257).

A minority of patients with pain have readily identifiable lesions whichare relatively easy to treat surgically or endoscopically. In otherpatients, pain is often thought to result from a variety of causes,including elevated intrapancreatic pressure, ischemia, and fibrosis.Without being bound by theory, however, these phenomena are not likelythe underlying cause of the pain. Rather, pain may result from abackground of neuronal sensitization induced by damage to theperineurium and subsequent exposure of the nerves to mediators andproducts of inflammation.

Given the importance of effective pain management in patients withchronic pancreatitis, additional therapies for treating painful symptomsare important and useful. TRPV3 inhibitors can be used to manage thepain associated with chronic pancreatitis. TRPV3 inhibitors can be usedalone or as part of an overall therapeutic treatment plan to managepatients with chronic pancreatits. For example, TRPV3 inhibitors can beadministered with pancreatic enzymes and/or insulin as part of atherapeutic regimen designed to manage patients with chronicpancreatitis.

Oral pain is a particular category of pain that may be treated using theTRPV3 inhibitors of the present invention. The term “oral pain” refersto any pain in the mouth, throat, lips, gums, teeth, tongue, or jaw. Theterm is used regardless of the cause of the pain and regardless ofwhether the oral pain is a primary or secondary symptom of a particulardisease, injury, or condition.

Oral pain has a large number of possible causes. In certain embodiments,oral pain is caused by an injury or disease of the mouth, jaw, teeth,gums, throat, lips, or tongue. In certain other embodiments, oral painis a consequence of an injury or disease that primarily affects anotherpart of the body. In still other embodiments, oral pain is a side effectof a therapy used to treat an injury or disease of the mouth or anotherpart of the body. TRPV3 inhibitors are useful in treating oral painregardless of its cause.

All pain has a serious negative impact on the health and well being ofthe sufferer. However, oral pain may have a particularly deleteriousimpact on patient health and quality of life. In particular, oral paincan interfere with appropriate eating and drinking. Thus, individualswith oral pain are susceptible to weight loss, malnutrition, anddehydration. In some instances, oral pain may interfere with hydrationand nutrition so significantly as to require intravenous, nasogastric,or other artificial support (e.g., tube feeding and/or hydration).Additionally, oral pain can interfere with proper oral hygiene. Poororal hygiene may further exacerbate many of the causes of oral pain, forexample, oral pain due to infection or abscess.

In certain embodiments, oral pain is caused by ulcers, sores, or otherlesions in the mouth. For example, oral pain may be caused by ulcers,sores, or other lesions on the tongue, gums, lips, throat, or othertissues of the mouth. Alternatively or additionally, oral pain may becaused by inflammation of the throat, tongue, gums, lips, or othertissues of the mouth. Inflammation may accompany ulcers or otherlesions, or inflammation may occur prior to or in the absence offormation of ulcers or other lesions.

The invention contemplates treatment of oral pain by administering aTRPV3 inhibitor by any route of administration described herein. Incertain embodiments, TRPV3 inhibitors for use in the treatment of oralpain are administered orally. Preferred preparations for oraladministration of TRPV3 inhibitors for use in treating oral pain are asa mouthwash, a gel, a tooth paste or other paste, a liquid, a lozenge,via a swab, or in association with a mouth guard or dental apparatus.The preparation and particular method of administration will depend onthe cause of the oral pain, the overall health and underlying medicalconditions of the patient, the severity of the pain, and othermedications or therapies the patient is concurrently receiving. Amedical practitioner can readily determine the optimal formulation foruse in a particular patient.

The conditions provided below are intended to illustrate the range ofinjuries and diseases of diverse etiology that may lead to oral pain.The invention contemplates administration of a TRPV3 inhibitor,according to the present invention, to treat or prevent oral pain. Incertain embodiments, compounds of the invention can be orallyadministered, for example as a gel, paste, mouth wash, or other oralpreparation, to help treat or prevent oral pain associated with anyinjury, disease, or condition. Regardless of the particular formulation,the invention contemplates administration by, for example, directapplication to the affected area of the mouth, rinsing of the entiremouth, via a swab, via a syringe, or on a mouth guard or other dentalapparatus.

For any of these conditions, the invention contemplates administrationof a TRPV3 inhibitor alone, or in combination with one or more othercompounds or treatment regimens appropriate for the particular injury orcondition.

Oral Mucositis

Oral mucositis, also known as stomatitis, is a common complication ofmany cancer treatments. Patients receiving systemic chemotherapy and/orlocal radiotherapy often develop extremely painful ulcers of the oralmucosa. This side effect is not limited to patients suffering fromcancers of the head and neck, but rather is a debilitating side effectafflicting approximately 40% of all chemotherapy patients (Preventionand Treatment of Oral Mucositis in Cancer Patients, 1998, Best Practice:2, pages 1-6.)

Oral mucositis is extremely painful. Additionally, oral mucositisinterferes with proper nutrition and hydration of cancer patients. Giventhe already compromised status of patients undergoing chemotherapyand/or radiotherapy, further interference with nutrition and hydrationmay seriously undermine patient health. Furthermore, these ulcerspresent an increased risk of infection. This risk is particularly acutein patients with compromised immune systems. Examples of patients atparticular risk of developing an opportunistic infection are patientswhose treatment included removal of one or more lymph nodes, patientswho previously received high-dose chemotherapy in preparation for a bonemarrow or stem cell transplant, and patients with an underlyingimmunosuppressive disorder (e.g., HIV or hepatitis).

Canker Sores

Canker sores, also known as aphthous ulcers (aphthae), may be relativelysmall and out-of-sight. However, they are often painful, persistent andannoying. Canker sores are shallow ulcers in the mouth that can makeeating and talking uncomfortable. They may occur on the tongue, softpalate, inside the cheek or lip, or at the base of the gums. Cankersores differ from cold sores in that they occur on the internal softtissues of the mouth and aren't contagious. Conversely, cold soresalmost always start out on the lips and don't often spread to the softtissues of the mouth. In addition, cold sores are caused by a form ofthe herpes virus, making them extremely contagious.

Researchers generally believe that stress or tissue injury may cause theeruption of canker sores. In some cases a minor injury, for examplebiting the inside of the mouth or eating rough foods, may trigger acanker sore. Other causes may include: (i) faulty immune systemfunction; (ii) nutritional problems, such as a deficiency of vitaminB-12, zinc, folic acid or iron; (iii) diseases of the gastrointestinaltract; (iv) food allergies; or (v) the menstrual cycle.

Canker sores can occur at any age, but often they first appear betweenthe ages of 10 and 40 years. Although canker sores typically resolve ontheir own, they can be very uncomfortable.

Dental/Tooth Abscess

Infection or decay can lead to an abscess. An abscess may have seriousdental and medical consequences. For example, a severe infection causedby a dental abscess may lead to a sinus or systemic infection.Furthermore, an abscess may lead to the need to extract one or moreteeth. Extraction may be necessary due to significant tooth decay, orbecause the infection is too severe to fully treat in the presence ofthe offending tooth.

Regardless of the ultimate outcome, a dental abscess may be extremelypainful. Not only is the pain uncomfortable, but it may interfere withproper nutrition and hydration. Methods and compositions for reducingthe pain associated with dental abscess would provide significantbenefits for their management.

Gastroesophageal Reflux Disease

Gastroesophageal reflux disease, or GERD, occurs when the loweresophageal sphincter (LES) does not close properly and stomach contentsleak back into the esophagus. The LES is a ring of muscle at the bottomof the esophagus that acts like a valve between the esophagus andstomach. When refluxed stomach acid touches the lining of the esophagus,it causes a burning sensation in the chest or throat. This is oftenexperienced as heartburn. The refluxed fluid may even be tasted in theback of the mouth, a sensation commonly referred to as acid indigestion.

Although occasional heartburn is uncommon and not necessarily indicativeof GERD, heartburn that occurs more than twice a week may be a sign ofGERD. In addition to the discomfort of heartburn and indigestion, GERDmay lead to other serious health problems. For example, over time, acidrefluxed to the back of the throat can lead to oral sores, lesions, orulcers in the mouth, gums, tongue, throat, or lips. The lesions cancause significant pain, can interfere with nutrition and hydration, andcan leave a person vulnerable to infection.

Administration of TRPV3 inhibitors, according to the present invention,may be useful in treating oral pain from lesions caused by GERD. TRPV3inhibitors may be used as part of a treatment regimen where the TRPV3inhibitor is administered to help manage the discomfort of the orallesion, while other agents or therapeutics interventions are used tomanage the GERD.

Gingivostomatitis

Gingivostomatitis is a disorder involving sores on the mouth and gumsthat result from a viral infection. Gingivostomatitis is characterizedby inflammation of the gums and mucosa and multiple oral ulcers. Theinflammation and ulcers are caused by viral infections, particularlythose that cause common childhood illness such as herpes virus (coldsores and acute herpetic stomatitis), and Coxsackie viruses (hand, footand mouth disease and herpangina). These viruses cause shallow ulcerswith a grayish or yellowish base and a slightly red margin, on thetissues of the gums (gingiva), the lining of the cheeks (buccal mucosa),or other soft tissues of the mouth. Although this condition can occur inpatients of any age, it is particularly common in children.

The oral ulcers casued by these viruses can be very painful. The ulcersare often accompanied by a fever. Overall, the condition can takeseveral weeks to resolve. The recognized treatments forgingivostomatitis focus on reducing the pain caused by the oral ulcers.This is particularly important for children who may refuse food orliquids because of their discomfort, thus making them especiallysusceptible to dehydration. Compounds of the present invention can beused to treat the pain associated with these oral ulcers.

Oral Thrush

Oral thrush is a fungal infection generally caused by the yeast fungus,Candida albicans, in the mucous membranes of the mouth. Strictlyspeaking, thrush is only a temporary Candida infection in the oralcavity of babies. However, the term is used generally to refer to fungalinfections in the mouths and throats of children and adults.

Candida is present in the oral cavity of almost half of the population.For example, everyone who wears dentures has Candida, withoutnecessarily suffering any ill effects. Generally, Candida does notcreate problems until there is a change in the chemistry of the oralcavity such that the growth of Candida is favored over the othermicroorganisms that typically inhabit the mouth and throat. Changes inoral chemistry sufficient to permit the growth of Candida may occur as aside effect to taking antibiotics or chemotherapeutics. Overall patienthealth may also influence the chemistry of the mouth. HIV infection,diabetes, malnutrition, age, and immunodeficiency are exemplaryconditions that can shift oral chemistry enough to permit the overgrowthof Candida in the mouth and throat.

In addition to shifts in oral chemistry, people whose dentures don't fitwell can sustain breaks in the mucous membranes in their mouth. Thesebreaks provide an opportunity for Candida infection in the mouth andlips.

Thrush causes white, cream-colored, or yellow spots in the mouth. Thespots are slightly raised. If these spots are scraped they tend tobleed. Thrush can be very uncomfortable, and may cause a burningsensation in the mouth and throat. The discomfort may interfere withhydration and nutrition. Furthermore, the discomfort may interfere withproper oral hygiene such as brushing and flossing.

Standard treatment of thrush is by administration of anti-fungal agents.These agents can be administered directly to the mouth, for example, inthe form of pastilles that are sucked or oral suspensions that are heldin the mouth before swallowing. Examples include nystatin (e.g., Nystanoral suspension), amphotericin (e.g., Fungilin lozenges) or miconazole(e.g., Daktarin oral gel). In addition to standard anti-fungal therapy,compounds of the present invention can be administered to manage thepain and discomfort associated with thrush.

Glossitis

Glossitis is an abnormality of the tongue that results frominflammation. Glossitis occurs when there is acute or chronicinflammation of the tongue. It causes the tongue to swell and changecolor. Finger-like projections on the surface of the tongue (papillae)are lost, causing the tongue to appear smooth. Glossitis has a number ofcauses including, but not limited to, the following: bacterialinfections; viral infections (including oral herpes simplex); injury ortrauma; exposure to irritants (e.g., tobacco, alcohol, hot foods,spices); allergic reactions; vitamin or mineral deficiencies (e.g., irondeficiency anemia, pernicious anemia and other B-vitamin deficiencies);or as a side effect of other diseases or disorders.

The symptoms of glossitis include swelling, soreness, and tenderness ofthe tongue. Additionally, the tongue often changes appearance, becomingsmooth and dark red in color. As a consequence of the swelling anddiscomfort, glossitis often makes chewing, swallowing, and speakingdiffcult.

The typical treatment for glossitis depends on the underlying cause ofthe inflammation. Regardless of the particular antibiotics,anti-inflammatories, or anti-viral agents that may be administered tocombat the underlying cause of glossitis, compounds according to thepresent invention may be administered to decrease the pain anddiscomfort associated with glossitis. Decreasing the pain associatedwith glossitis is especially important when it interferes with propernutrition and hydration, or when it interferes with or prevents properoral hygiene.

Cutaneous Diseases

Oral ulcers may result from any of a number of cutaneous diseases. Forexample, lichen planus, pemphigus, pemphigoid, and erythema multiformemay lead to oral ulcers. Such oral ulcers may cause significant painthat can be treated using the compounds of the present invention.

Reduction of pain may help facilitate healing. This is especiallyimportant for patients with pemphigus and pemphigoid who develop oralulcers. Such patients are already immunosuppressed, and may thus be moresusceptible to opportunistic infections from lesions in the mouth.

Gastrointestinal Diseases

Oral ulcers may result from any of a number of gastrointestinaldiseases. Conditions which interfere with proper digestion, managementand flow of stomach and other digestive acids, motility, and eliminationmay lead to oral ulcers and other lesions. In some instances, the oralulcers are the results of acids or partially digested food refluxinginto the esophagus. In other instances, the oral ulcers result fromfrequent vomiting. In still other instances, oral ulcers occur due tovitamin deficiency, mineral deficiency or other nutritional deficiencysecondary to the gastrointestinal disease. In still other instances,oral ulcers are part of the complex etiology that characterizes thegastrointestinal disease.

Oral ulcers resulting from or experienced as part of a gastrointestinaldisease may be extremely painful. They may undermine proper nutritionand hydration for a patient whose underlying gastrointestinal diseasemay already impose multiple limitations on diet. Accordingly, methodsand compositions for decreasing the discomfort and pain associated withthese oral ulcers offer substantial benefits for patients with anunderlying gastrointestinal condition.

Exemplary gastrointestinal conditions which may lead to oralinflammation, lesions, or ulcers include, but are not limited to,Crohn's disease, ulcerative colitis, irritable bowel syndrome, celiacsprue, and dermatitis herpetiformis. The primary symptoms of theseconditions may be managed with diet, stress management, and medications.The TRPV3 inhibitors of the present invention may be used to help managethe pain and discomfort of oral inflammation, lesions, or ulcers causedby any of these gastrointestinal conditions.

Rheumatoid Diseases

A consequence of several rheumatoid diseases is oral ulcers. Forexample, lupus, Behcet's syndrome, Sweet's syndrome, and Reiter'sdisease may all lead to oral ulcers. Such oral ulcers may causesignificant mouth pain that can be treated using the compounds of thepresent invention.

Sjogren's Syndrome

Dry mouth is a common symptom associated with S{umlaut over (j)}ogren'ssyndrome. Dry mouth is caused by a decrease in the production of saliva.Saliva is an essential body fluid for protection and preservation of theoral cavity and oral functions. Although saliva is mostly water, it alsocontains over 60 substances which serve the following importantfunctions: protect, lubricate and cleanse the oral mucosa; aid chewing,swallowing and talking; protect the teeth against decay; protect themouth, teeth, and throat from infection by bacteria, yeasts, andviruses; support and facilitate our sense of taste.

Given the important functions of saliva, decreased salivation can leadto many problems. If the condition persists for months or years, apatient may develop oral complications such as difficulty swallowing,severe and progressive tooth decay, oral infections (particularlyfungal), or combinations of these. Many of the conditions can causediscomfort, in their own right, and may also lead to oral lesions orulcers.

Several medications are available to help increase salivary secretion inpatients with dry mouth. Pilocarpine (Salagen®) and cevimeline (Evoxac®)reduce symptoms of dry mouth and increase salivary secretion. However,these drugs don't prevent tooth decay or treat the oral pain associatedwith the symptoms or effects of dry mouth. Compounds of the presentinvention can be used to treat the pain associated with dry mouth.

Vitamin or Mineral Deficiencies

In some instances, vitamin or mineral deficiencies may lead to ulcers orother sores in the mouth. For example, deficiency in vitamin C may leadto the oral lesions characteristic of scurvy. Deficiencies in vitaminsB1, B2, B6, or B12 may also lead to oral lesions. Additionally,deficiencies in zinc, folic acid, iron, selenium, or calcium may lead tooral lesions.

In certain embodiments, a vitamin or mineral deficiency is aprecipitating factor leading to a canker sore. However, a vitamin ormineral deficiency may also lead to other types of oral ulcers andlesions. Regardless of the nature of the lesion, compounds of thepresent invention can be used to help manage the associated pain.

Allergies

Allergies can sometimes lead to canker sores and other oral lesions.Oral lesions due to an allergy may be more likely when a person's oraltissues come into contact with the causative allergen. However, contactbetween the allergen and oral tissue is not necessarily required toproduce an oral lesion. Exemplary allergens that can lead to orallesions include food allergens such as fruits and vegetables (e.g.,strawberries, lemons, oranges, pineapples, apples, figs, tomatoes);shellfish; chocolate; nuts; dairy (e.g., milk and cheese); cereal grains(e.g., buckwheat, wheat, oats, rye, barley, gluten protein found ingrains); additives (e.g., cinnamonaldehyde (a flavoring agent), benzoicacid (a preservative); toothpastes (e.g., some people have a sensitivityto sodium laurel sulfate found in certain toothpastes and mouthwashes);nonsteroidal anti-inflammatory drugs (NSAIDs; some people have asensitivity leading to canker sores in response to this class of drug).

Other Exemplary Conditions and Injuries

The foregoing are merely exemplary of diseases and conditions that causeor lead to inflammation, lesions, ulcers, or other sources of oral pain.In other embodiments, the oral pain is due to an injury to the mouth,jaw, lips, gums, or teeth. In other embodiments, the oral pain is due tooral surgery, for example, surgery for cancer, tooth extraction, or jawremodeling. Other conditions that may lead to oral ulcers, and thus oralpain, include, but are not limited to chickpox, herpes zoster,infectious mononucleosis, syphilis, tuberculosis, acute necrotizinggingivitis, and burning mouth syndrome. Additionally, conditions thatlead to a compromised immune system put patients at risk for, amongother complications, oral inflammation, lesions, or ulcers. HIVinfection, AIDS, and hepatitis are all conditions that undermine theimmune system and may lead to oral lesions or ulcers. Additionally,individuals taking immunosuppressants (e.g., organ transplantrecipients, bone marrow recipients, stem cells recipients, patients withan autoimmune disease) are at increased risk of developing painful orallesions.

The invention contemplates the use of TRPV3 inhibitors, according to thepresent invention, in the treatment of oral pain regardless of theunderlying cause. In certain embodiments, TRPV3 inhibitors for treatingoral pain can be administered orally, for example, as a paste, mouthwash, gel, or other liquid preparation. In certain embodiments, thepaste, mouth wash, gel, or other liquid preparation is administered viaa swab, mouth guard, or other dental apparatus. In certain embodiments,the preparation is applied locally to the mouth, but is not otherwiseingested. For example, a mouth wash formulation that is not swallowedmay be used. Regardless of the formulaion and route of administeration,the invention contemplates administration of the subject TRPV3inhibitors as part of an overall treatment strategy that also includestherapies appropriate for the particular disease or condition thatcaused the oral inflammation, lesion, or ulcer.

TRPV3 inhibitors may be used to treat oral pain resulting from any ofthe foregoing injuries, diseases, or conditions. Additionally,Applicants note that the subject TRPV3 inhibitors may also be useful inthe treatment of the underlying aforementioned diseases and conditionsthemselves. Specifically, TRPV3 inhibitors may be useful in thetreatment of inflammation, and thus diseases or conditions with aninflammatory component, whether the symptoms manifest themselves in themouth or in other parts of the body, may themselves be treatable withthe subject TRPV3 inhibitors. Accordingly, the invention contemplatesand recognizes that for some conditions the therapeutic affects ofadministering a TRPV3 inhibitor may be two-fold: (i) decreasing painassociated with one or more symptoms of a disease or condition and (ii)treating the underlying symptoms or disease.

Disease and Injury Models

Compounds that antagonize TRPV3 function may be useful in theprophylaxis and treatment of any of the foregoing injuries, diseases,disorders, or conditions. In addition to in vitro assays of the activityof these compounds, their efficacy can be readily tested in one or moreanimal models. By way of example, numerous well known animal modelsexist. One or more suitable animal models (e.g., suitable in light ofthe particular indication) can be selected.

Pain can be generally categorized as chronic pain and acute pain. Thetwo categories of pain differ in duration, as well as underlyingmechanism. Chronic pain is not only persistent, but also does notgenerally respond well to treatment with currently available analgesics,non-steroidal anti-inflammatory drugs, and opioids.

Two broad sub-categories of chronic pain are neuropathic pain and cancerpain. Wang and Wang (2003) Advanced Drug Delivery Reviews 55: 949-965.Neuropathic pain refers to pain resulting from damage (e.g., fromdisease, injury, age) to the nervous system (e.g., nerves, spinal cord,CNS, PNS). Cancer-related pain may be caused by tumor infiltration,nerve compression, substances secreted by tumors, or the particulartreatment regimen (e.g., radiation, chemotherapeutics, surgery).

Pain is also often classified mechanistically as nociceptive,inflammatory, or neuropathic. Nociceptive pain is pain experiencedfollowing, for example, changes or extremes in temperature, exposure toacids, exposure to chemical agents, exposure to force, and exposure topressure. Reception of painful stimuli sends impulses to the dorsal rootganglia. The response is typically a combination of a reflexive response(e.g., withdrawal from the stimuli) and an emotional reaction.Inflammation is the immune system's response to injury or disease. Inresponse to injury or disease, macrophages, mast cells, neutrophils, andother cells of the immune system are recruited. This infiltration ofcells, along with the release of cytokines and other factors (e.g.,histamine, serotonin, bradykinin, prostaglandins, ATP, H+, nerve growthfactor, TNFα, endothelins, interleukins), can cause fever, swelling, andpain. Current treatments for the pain of inflammation include Cox2inhibitors and opioids. Neuropathic pain refers to pain resulting fromdamage (e.g., from disease, injury, age) to the nervous system (e.g.,nerves, spinal cord, CNS, PNS). Current treatment for neuropathic painincludes tricyclic antidepressants, anticonvulsants, Na+ channelblockers, NMDA receptor antagonists, and opioids.

There are numerous animal models for studying pain. Generally, theanimal models mimic one of the foregoing mechanisms of pain, rather thanthe pain associated with any one disease or injury. Such models provideevidence of whether a drug or therapy would be effective in treating anyof a number of injuries, diseases, or conditions that generate pain viaa particular mechanism (e.g., nociceptive, inflammatory, orneuropathic).

The various models use various agents or procedures to simulate painresulting from injuries, diseases, or other conditions. Blackburn-Munro(2004) Trends in Pharmacological Sciences 25: 299-305 (see, for example,Table 1). Behavioral characteristics of challenged animals can then beobserved. Compounds or procedures that may reduce pain in the animalscan be readily tested by observing behavioral characteristics ofchallenged animals in the presence versus the absence of the testcompound(s) or procedure.

Exemplary behavioral tests used to study chronic pain include tests ofspontaneous pain, allodynia, and hyperalgesia. Id. To assess spontaneouspain, posture, gait, nocifensive signs (e.g., paw licking, excessivegrooming, excessive exploratory behavior, guarding of the injured bodypart, and self-mutilation) can be observed. To measure evoked pain,behavioral responses can be examined following exposure to heat (e.g.,thermal injury model).

Exemplary animal models of pain include, but are not limited to, theChung model, the carageenan induced hyperalgesia model, the Freund'scomplete adjuvant (CFA) induced hyperalgesia model, the thermal injurymodel, the formalin model and the Bennett Model. The Chung model ofneuropathic pain (without inflammation) involves ligating one or morespinal nerves. Chung et al. (2004) Methods Mol Med 99: 35-45; Kim andChung (1992) Pain 50: 355-363. Ligation of the spinal nerves results ina variety of behavioral changes in the animals including heathyperalgesia, cold allodynia, and ongoing pain. Compounds thatantagonize TRPV3 can be administered to ligated animals to assesswhether they diminish these ligation induced behavioral changes incomparison to that observed in the absence of compound.

Carageenan induced hyperalgesia and Freund's complete adjuvent (CFA)induced hyperalgesia are models of inflammatory pain. Walker et al.(2003) Journal of Pharmacol Exp Ther 304: 56-62; McGaraughty et al.(2003) Br J Pharmacol 140: 1381-1388; Honore et al. (2005) J PharmacolExp Ther. Compounds that antagonize TRPV3 can be administered tocarrageenan or FCA challenged animals to assess whether they diminishthermal hyperalgesia in comparison to that observed in the absence ofcompound. In addition, the ability of compounds that antagonize TRPV3function to diminish cold and/or mechanical hypersensitivity can also beassessed in these models. Typically, the carrageenan inducedhyperalgesia model is believed to mimic acute inflammatory pain and theCFA model is believed to mimic chronic pain and chronic inflammatorypain.

The Bennett model uses prolonged ischemia of the paw to mirror chronicpain. Xanthos et al. (2004) J Pain 5: S1. This provides an animal modelfor chronic pain including post-operative pain, complex regional painsyndrome, and reflex sympathetic dystrophy. Prolonged ischemia inducesbehavioral changes in the animals including hyperalgesia to mechanicalstimuli, sensitivity to cold, pain behaviors (e.g., paw shaking,licking, and/or favoring), and hyperpathia. Compounds that antagonizeTRPV3 can be administered to challenged animals to assess whether theydiminish any or all of these behaviors in comparison to that observed inthe absence of compound. Similar experiments can be conducted in athermal injury model which can be used to mimic post-operative pain.

Additional models of neuropathic pain include central pain models basedon spinal cord injury. Chronic pain is generated by inducing a spinalcord injury, for example, by dropping a weight on a surgically exposedarea of spinal cord (e.g., weight-drop model). Spinal cord injury canadditionally be induced by crushing or compressing the spinal cord, bydelivering neurotoxin, using photochemicals, or by hemisecting thespinal cord. Wang and Wang (2003).

Additional models of neuropathic pain include peripheral nerve injurymodels. The term peripheral neuropathy encompasses a variety ofdiseases, conditions, and injuries. One of skill in the art can readilyselect an appropriate model in light of the particular condition ordisease under investigation. Exemplary models include, but are notlimited to, the neuroma model, the Bennett model, the Seltzer model, theChung model (ligation at either L5 or L5/L6), the sciatic cryoneurolysismodel, the inferior caudal trunk resection model, and the sciaticinflammatory neuritis model. Id.

Exemplary models of neuropathic pain associated with particular diseasesare also available. Diabetes and shingles are two diseases oftenaccompanied by neuropathic pain. Even following an acute shinglesepisodes, some patients continue to suffer from postherpetic neuralgiaand experience persistent pain lasting years. Neuropathic pain caused byshingles and/or postherpetic neuralgia can be studied in thepostherpetic neuralgia model (PHN). Diabetic neuropathy can be studiedin diabetic mouse models, as well as chemically induced models ofdiabetic neuropathy. Wang and Wang (2003).

As outlined above, cancer pain may have any of a number of causes, andnumerous animal models exist to examine cancer pain related to, forexample, chemotherapeutics or tumor infiltration. Exemplary models oftoxin-related cancer pain include the vincristine-induced peripheralneuropathy model, the taxol-induced peripheral neuropathy model, and thecisplatin-induced peripheral neuropathy model. Wang and Wang (2003). Anexemplary model of cancer pain caused by tumor infiltration is thecancer invasion pain model (CIP). Id.

Primary and metastatic bone cancers are associated with tremendous pain.Several models of bone cancer pain exist including the mouse femur bonecancer pain model (FBC), the mouse calcaneus bone cancer pain model(CBC), and the rat tibia bone cancer model (TBC). Id.

An additional model of pain is the formalin model. Like the carrageenanand CFA models, the formalin model involves injection of an irritantintradermally or intraperitoneally into an animal. Injection offormalin, a 37 percent solution of formaldehyde, is the most commonlyused agent for intradermal paw injection (the formalin test). Injectionof a 0.5 to 15 percent solution of formalin (usually about 3.5%) intothe dorsal or plantar surface of the fore- or hindpaw produces abiphasic painful response of increasing and decreasing intensity forabout 60 minutes after the injection. Typical responses include the pawbeing lifted, licked, nibbled, or shaken. These responses are considerednociceptive. The initial phase of the response (also known as the EarlyPhase), which lasts 3 to 5 minutes, is probably due to direct chemicalstimulation of nociceptors. This is followed by 10 to 15 minutes duringwhich animals display little behavior suggestive of nociception. Thesecond phase of this response (also known as the Late Phase) startsabout 15 to 20 minutes after the formalin injection and lasts 20 to 40minutes, initially rising with both number and frequency of nociceptivebehaviors, reaching a peak, then falling off. The intensities of thesenociceptive behaviors are dependent on the concentration of formalinused The second phase involves a period of sensitization during whichinflammatory phenomena occur. The two phases of responsiveness toformalin injection makes the formalin model an appropriate model forstudying mociceptive and acute inflammatory pain. It may also model, insome respects, neuropathic pain.

In addition to any of the foregoing models of chronic pain, compoundsthat antagonize TRPV3 function can be tested in one or more models ofacute pain. Valenzano et al. (2005) Neuropharmacology 48: 658-672.Regardless of whether compounds are tested in models of chronic pain,acute pain, or both, these studies are typically (though notexclusively) conducted, for example, in mice, rats, or guinea pigs.Additionally, compounds can be tested in various cell lines that providein vitro assays of pain. Wang and Wang (2003).

The foregoing animal models are relied upon extensively in the study ofpain. The following provide additional exemplary references describingthe use of these models in the study of pain: thermal injury model(Jones and Sorkin, 1998, Brain Res 810: 93-99; Nozaki-Taguchi and Yaksh,1998, Neuroscience Lett 254: 25-28; Jun and Yaksh, 1998, Anesth Analg86: 348-354), formalin model (Yaksh et al., 2001, J Appl Physiol 90:2386-2402), carrageenan model (Hargreaves et al., 1988, Pain 32: 77-88),and CFA model (Nagakura et al., 2003, J Pharmacol Exp Ther 306:490-497).

For testing the efficacy of TRPV3 antagonists for the treatment ofcough, experiments using the conscious guinea pig model of cough can bereadily conducted. Tanaka and Maruyama (2003) Journal Pharmacol Sci 93:465-470; McLeod et al. (2001) Br J Pharmacol 132: 1175-1178. Briefly,guinea pigs serve as a useful animal model for cough because, unlikeother rodents such as mice and rats, guinea pigs actually cough.Furthermore, guinea pig coughing appears to mimic human coughing interms of the posture, behavior, and appearance of the coughing animal.

To induce cough, conscious guinea pigs are exposed to an inducing agentsuch as citric acid or capsaicin. The response of the animal is measuredby counting the number of coughs. The effectiveness of a coughsuppressing agent, for example a compound that inhibits TRPV3, can bemeasured by administering the agent and assessing the ability of theagent to decrease the number of coughs elicited by exposure to citricacid, capsaicin, or other similar cough-inducing agent. In this way,TRPV3 inhibitors for use in the treatment of cough can be readilyevaluated and identified.

Additional models of cough include the unconscious guinea pig model.Rouget et al. (2004) Br J Pharmacol 141: 1077-1083. Either of theforegoing models can be adapted for use with other animals capable ofcoughing. Exemplary additional animals capable of coughing include catsand dogs.

As detailed above, TRPV3 inhibitors can be used to treat the symptoms ofpain associated with pancreatitis. The efficacy of TRPV3 inhibitors inpancreatitis pain management may be tested in one or more animal modelsInhibitors may be tested in general animal models of pain, for examplemodels of inflammatory pain or visceral pain. Alternatively oradditionally, TRPV3 inhibitors may be tested in animal models thatspecifically mimic pain accompanying pancreatitis or other pancreaticinjury.

Several rat models of pancreatitic pain have recently been described(Lu, 2003, Anesthesiology 98(3): 734-740; Winston et al., 2003, Journalof Pain 4(6): 329-337). Lu et al. induced pancreatitis by systemicdelivery of dibutylin dichloride in rats. Rats showed an increase inwithdrawal events after von Frey filament stimulation of the abdomen anddecreased withdrawal latency after thermal stimulation during a periodof 7 days. The pain state induced in these animals was alsocharacterized by increased levels of substance P in spinal cords (Lu, etal., 2003). To test the efficacy of a TRPV3 inhibitor in this model, aTRPV3 inhibitor can be administered following or concurrently withdelivery of dibutylin dichloride. Control animals can be administered acarrier or a known pain reliever. Indicia of pain can be measured.Efficacy of a TRPV3 inhibitor can be evaluated by comparing the indiciaof pain observed in animals receiving a TRPV3 inhibitor to that ofanimals that did not receive a TRPV3 inhibitor. Additionally, efficacyof a TRPV3 inhibitor can be compared to that of known pain medicaments.

The efficacy of von Frey filament testing as a means to measurenociceptive behavior was also shown by inducing pancreatitis by systemicL-arginine administration (Winston et al, 2003). The efficacy of a TRPV3inhibitor can similarly be tested following pancreatitis induced bysystemic L-arginine administration.

Lu et al. also described direct behavioral assays for pancreatic painusing acute noxious stimulation of the pancreas via an indwelling ductalcanula in awake and freely moving rats. These assays included cagecrossing, rearing, and hind limb extension in response tointrapancreatic bradykinin infusion. Intrathecal administration ofeither D-APV (NMDA receptor antagonist) or morphine alone partiallyreduced visceral pain behaviors in this model. Combinations of bothreduced pain behaviors to baseline. The efficacy of a TRPV3 inhibitorcan similarly be tested in this system.

Any of the foregoing animal models may be used to evaluate the efficacyof a TRPV3 inhibitor in treating pain associated with pancreatitis. Theefficacy can be compared to a no teatment or placebo control.Additionally or alternatively, efficacy can be evaluated in comparisonto one or more known pain relieving medicaments.

Optimizing the Treatment of Pain

TRPV3 inhibitors, according to the present invention, can be used in thetreatment of a variety of injuries, diseases, conditions, and disorders.One important therapeutic use for TRPV3 inhibitors is in the treatmentof pain. As illustrated by the extensive list of injuries, conditions,and diseases for which pain is a significant and sometimes debilitatingsymptom, improved methods and compositions for use in the treatment ofpain provide substantial benefits for an enormous range of patients.Such methods and compositions have the potential to improve the qualityof care and the quality of life for patients afflicted with a diverserange of injuries, diseases, and conditions.

TRPV3 is a good target for modulating pain. TRPV3 is expressed intissues that contribute to transmission of painful stimuli.Additionally, TRPV3 expression is upregulated, for example in dorsalroot ganglia, following injury. Finally, TRPV3 knockout mice exhibitabnormal responses to painful stimuli. These characteristics of TRPV3suggest that inhibitors of TRPV3 will be useful in the treatment ofpain.

Many of these characteristics are shared by TRPV1, and inhibitors ofTRPV1 are being developed for the treatment of pain. However, althoughTRPV1 and TRPV3 share certain characteristics consistent with thedevelopment of effective therapeutics for the treatment of pain, TRPV3possesses certain characteristics that makes it a better target fortherapeutic compounds for the treatment of pain. For example, TRPV3sensitizes upon repeated stimulation. In contrast, TRPV1 desensitizesupon repeated stimulation with the agonist capsaicin. In addition toexpression in dorsal root ganglia, TRPV3 is expressed in skin. Given thesignificant involvement of skin in many types of pain, this expressionpattern is suggestive of potential effectiveness of TRPV3 inhibitors inpain involving the skin.

An important issue with the treatment of pain is how to manage painwhile reducing the side effects experienced with many analgesics. Forexample, although many opiates and other narcotics effectively diminishpain, patients are often unable to drive, work, or concentrate whiletaking these medications. Thus, while opiates such as morphine ordilaudin may be suitable for short term use or for use duringhospitalization, they are not optimal for long term use. Additionally,opiates and other narcotics are habit forming, and patients typicallydevelop a tolerance for these drugs. These characteristics of opioidsand other narcotics make them sub-optimal for pain management.

The present invention provides TRPV3 inhibitors for use in vitro and invivo. The present invention also provides compositions andpharmaceutical compositions comprising particular classes of compoundsthat inhibit TRPV3 activity. In certain embodiments, the subject TRPV3inhibitors are selective. In other words, in certain embodiments, thecompound inhibits TRPV3 activity preferentially over the activity ofother ion channels. In certain embodiments, the compound inhibits TRPV3activity preferentially over TRPV1, TRPV2, TRPV4, and/or TRPM8 activity.In certain other embodiments, the compound is selected because it crossreacts with one or more other TRP channels involved with pain. Forexample, in certain embodiments, the compound inhibits the activity ofTRPV3 and also inhibits the activity of one or more of TRPV1, TRPV2,TRPV4, and TRPM8.

Combination Therapy

Another aspect of the invention provides a conjoint therapy wherein oneor more other therapeutic agents are administered with the TRPV3modulators. Such conjoint treatment may be achieved by way of thesimultaneous, sequential, or separate dosing of the individualcomponents of the treatment.

In certain embodiments, a compound of the invention is conjointlyadministered with an analgesic. Suitable analgesics include, but are notlimited to, opioids, glucocorticosteroids, non-steroidalanti-inflammatories, naphthylalkanones, oxicams, para-aminophenolderivatives, propionic acids, propionic acid derivatives, salicylates,fenamates, fenamate derivatives, pyrozoles, and pyrozole derivatives.Examples of such analgesic compounds include, but are not limited to,codeine, hydrocodone, hydromorphone, levorpharnol, morphine, oxycodone,oxymorphone, butorphanol, dezocine, nalbuphine, pentazocine, etodolac,indomethacin, sulindac, tolmetin, nabumetone, piroxicam, acetaminophen,fenoprofen, flurbiprofen, ibuprofen, ketoprofen, naproxen, diclofenac,oxaprozin, aspirin, diflunisal, meclofenamic acid, mefanamic acid,prednisolone, and dexamethasone. Preferred analgesics are non-steroidalanti-inflammatories and opioids (preferably morphine).

In certain embodiments, a compound of the invention is conjointlyadministered with a non-steroidal anti-inflammatory. Suitablenon-steroidal anti-inflammatory compounds include, but are not limitedto, piroxicam, diclofenac, etodolac, indomethacin, ketoralac, oxaprozin,tolmetin, naproxen, flubiprofen, fenoprofen, ketoprofen, ibuprofen,mefenamic acid, sulindac, apazone, phenylbutazone, aspirin, celecoxiband rofecoxib.

In certain embodiments, a compound of the invention is conjointlyadministered with an antiviral agent. Suitable antiviral agents include,but are not limited to, amantadine, acyclovir, cidofovir, desciclovir,deoxyacyclovir, famciclovir, foscamet, ganciclovir, penciclovir,azidouridine, anasmycin, amantadine, bromovinyldeoxusidine,chlorovinyldeoxusidine, cytarbine, didanosine, deoxynojirimycin,dideoxycitidine, dideoxyinosine, dideoxynucleoside, edoxuidine,enviroxime, fiacitabine, foscamet, fialuridine, fluorothymidine,floxuridine, hypericin, interferon, interleukin, isethionate,nevirapine, pentamidine, ribavirin, rimantadine, stavirdine,sargramostin, suramin, trichosanthin, tribromothymidine,trichlorothymidine, vidarabine, zidoviridine, zalcitabine3-azido-3-deoxythymidine, 2′,3′-dideoxyadenosine (ddA),2′,3′-dideoxyguanosine (ddG), 2′,3′-dideoxycytidine (ddC),2′,3′-dideoxythymidine (ddT), 2′3′-dideoxy-dideoxythymidine (d4T),2′-deoxy-3′-thia-cytosine (3TC or lamivudime),2′,3′-dideoxy-2′-fluoroadenosine, 2′,3′-dideoxy-2′-fluoroinosine,2′,3′-dideoxy-2′-fluorothymidine, 2′,3′-dideoxy-2′-fluorocytosine,2′3′-dideoxy-2′,3′-didehydro-2′-fluorothymidine (Fd4T),2′3′-dideoxy-2′-beta-fluoroadenosine (F-ddA),2′3′-dideoxy-2′-beta-fluoro-inosine (F-ddI), and2′,3′-dideoxy-2′-beta-flurocytosine (F-ddC), trisodiumphosphomonoformate, trifluorothymidine, 3′azido-3′thymidine (AZT),dideoxyinosine (ddI), and idoxuridine.

In certain embodiments, a compound of the invention is conjointlyadministered with an antibacterial agent. Suitable antibacterial agentsinclude, but are not limited to, amanfadine hydrochloride, amanfadinesulfate, amikacin, amikacin sulfate, amoglycosides, amoxicillin,ampicillin, amsamycins, bacitracin, beta-lactams, candicidin,capreomycin, carbenicillin, cephalexin, cephaloridine, cephalothin,cefazolin, cephapirin, cephradine, cephaloglycin, chilomphenicols,chlorhexidine, chloshexidine gluconate, chlorhexidine hydrochloride,chloroxine, chlorquiraldol, chlortetracycline, chlortetracyclinehydrochloride, ciprofloxacin, circulin, clindamycin, clindamycinhydrochloride, clotrimazole, cloxacillin, demeclocycline,diclosxacillin, diiodohydroxyquin, doxycycline, ethambutol, ethambutolhydrochloride, erythromycin, erythromycin estolate, erhmycin stearate,farnesol, floxacillin, gentamicin, gentamicin sulfate, gramicidin,giseofulvin, haloprogin, haloquinol, hexachlorophene, iminocylcline,iodochlorhydroxyquin, kanamycin, kanamycin sulfate, lincomycin,lineomycin, lineomycin hydrochoride, macrolides, meclocycline,methacycline, methacycline hydrochloride, methenine, methenaminehippurate, methenamine mandelate, methicillin, metonidazole, miconazole,miconazole hydrochloride, minocycline, minocycline hydrochloride,mupirocin, nafcillin, neomycin, neomycin sulfate, netimicin, netilmicinsulfate, nitrofurazone, norfloxacin, nystatin, octopirox, oleandomycin,orcephalosporins, oxacillin, oxyteacline, oxytetracycline hydrochloride,parachlorometa xylenol, paromomycin, paromomycin sulfate, penicillins,penicillin G, penicillin V, pentamidine, pentamidine hydrochloride,phenethicillin, polymyxins, quinolones, streptomycin sulfate,tetracycline, tobramycin, tolnaftate, triclosan, trifampin, rifamycin,rolitetracycline, spectinomycin, spiramycin, struptomycin, sulfonamide,tetracyclines, tetracycline, tobramycin, tobramycin sulfate,triclocarbon, triclosan, trimethoprim-sulfamethoxazole, tylosin,vancomycin, and yrothricin.

In certain embodiments, a compound of the invention is conjointlyadministered with a cough suppressant, decongestant, or expectorant.

Examples of retinoids that be administered with the subject TRPV3inhibitors, e.g., where the TRPV3 inhibitor can be used to reduce thepain and/or inflammatory effect of the retinoid, include, but are notlimited to, compounds such as retinoic acid (both cis and trans),retinol, adapalene, vitamin A and tazarotene. Retinoids are useful intreating acne, psoriasis, rosacea, wrinkles and skin cancers and cancerprecursors such as melanoma and actinic keratosis.

Similarly, the subject TRPV3 inhibitors can be used in conjunction withkeratolytic agents include benzoyl peroxide, alpha hydroxyacids, fruitacids, glycolic acid, salicylic acid, azelaic acid, trichloroaceticacid, lactic acid and piroctone.

The subject TRPV3 inhibitors can also be administered along withdepilatory agents (hair loss).

The subject TRPV3 inhibitors can be used with anti-acne agents,anti-eczema agents and anti-psoratic agents. Compounds particlarlyuseful in treating acne include azelaic acid (an aliphatic diacid withantiacne properties), anthralin (a diphenolic compound with antifungaland antipsoriatic properties), and masoprocol (nordihydroguaiareticacid, a tetraphenolic compound with antioxidant properties, also usefulin the treatment of actinic keratosis) and analogs thereof (such asaustrobailignan 6, oxoaustrobailignan 6,4′-O-methyl-7,7′-dioxoaustrobailignan 6, macelignan,demethyldihydroguaiaretic acid, 3,3′,4-trihydroxy-4′-methoxylignan,Saururenin, 4-hydroxy-3,3′,4′-trimethoxylignan, and isoanwulignan).Anti-eczema agents include pimecrolimus and tacrolimus. Anti-psoriaticactive agents suitable for use in the present invention includeretinoids (including isomers and derivatives of retinoic acid, as wellas other compounds that bind to the retinoic acid receptor, such asretinoic acid, acitretin, 13-cis-retinoic acid (isotretinoin),9-cis-retinoic acid, tocopheryl-retinoate (tocopherol ester of retinoicacid (trans- or cis-)), etretinate, motretinide,1-(13-cis-retinoyloxy)-2-propanone,1-(13-cis-retinoyloxy)-3-decanoyloxy-2-propanone,1,3-bis-(13-cis-retinoyloxy)-2-propanone,2-(13-cis-retinoyloxy)-acetophenone,13-cis-retinoyloxymethyl-2,2-dimethyl propanoate,2-(13-cis-retinoyloxy)-n-methyl-acetamide,1-(13-cis-retinoyloxy)-3-hydroxy-2-propanone,1-(13-cis-retinoyloxy)-2,3-dioleoylpropanone, succinimdyl13-cis-retinoate, adapalene, and tazarotene), salicylic acid(monoammonium salt), anthralin, 6-azauridine, vitamin D derivatives(including but not limited to Rocaltrol (Roche Laboratories), EB 1089(24α,26α,27α-trihomo-22,24-diene-1α, 25-(OH)₂-D₃), KH 1060(20-epi-22-oxa-24α,26α,27α-trihomo-1α,25-(OH)₂-D₃), MC 1288, GS 1558, CB1093, 1,25-(OH)₂-16-ene-D₃, 1,25-(OH)₂-16-ene-23-yne-D₃, and25-(OH)2-16-ene-23-yne-D₃, 22-oxacalcitriol; 1α-(OH)D₅ (University ofIllinois), ZK 161422 and ZK 157202 (Institute of MedicalChemistry-Schering AG), alfacalcidol, calcifediol, calcipotriol(calcipotriene), maxacalcitriol, colecalciferol, doxercalciferol,ergocalciferol, falecalcitriol, lexacalcitol, maxacalcitol,paricalcitol, secalciferol, seocalcitol, tacalcitol, calcipotriene,calcitriol, and other analogs as disclosed in U.S. Pat. No. 5,994,332),pyrogallol, and tacalcitol.

The subject TRPV3 inhibitors can also be administered with vitamins andderivatives thereof including Vitamin A, ascorbic acid (Vitamin C),alpha-tocopherol (Vitamin E), 7-dehydrocholesterol (Vitamin D), VitaminK, alpha-lipoic acid, lipid soluble anti-oxidants, and the like.

The subject TRPV3 inhibitors can also be used with skin protectants,such allantoin and esculin.

In certain embodiments, two or more compounds of the invention areconjointly administered. When two or more compounds of the invention areconjointly administered, the two or more compounds may have a similarselectivity profile and functional activity, or the two or morecompounds may have a different selectivity profile and functionalactivity. By way of example, the two or more compounds may both beapproximately 10, 100, or 1000 fold selective for antagonizing afunction of TRPV3 over TRPV1, TRPV5, and TRPV6 (e.g., the two or morecompounds have a similar selectivity profile), and further may inhibit afunction of TRPV3 with a similar IC₅₀ (e.g., a similar functionalactivity). Alternatively, the one of the two or more compounds mayselectively inhibit TRPV3 while the other of the two or more compoundsinhibits both TRPV3 and TRPV1 (e.g., the two or more compounds havediffering selectivity profiles). Administration of combinations of twoor more compounds of the invention having similar or differingproperties are contemplated.

In certain embodiments, a compound of the invention is conjointlyadministered with one or more additional compounds that antagonize thefunction of a different channel. By way of example, a compound of theinvention may be conjointly administered with one or more compounds thatantagonize TRPV1 and/or TRPV4. The compound(s) that antagonize TRPV1 orTRPV4 may be selective for TRPV1 or TRPV4 (e.g, inhibit TRPV1 or TRPV410, 100, or 1000 fold more strongly than TRPV3). Alternatively, thecompound(s) that antagonize TRPV1 or TRPV4 may cross react with otherTRP channels.

In certain other embodiments, a compound of the invention is conjointlyadministered with one or more additional agents or therapeutic regimensappropriate for the particular injury, disease, condition, or disorderbeing treated.

When combinations of a TRPV3 inhibitor and one or more other compounds,agents, or therapeutic regimens are administered, the inventioncontemplates administration via the same route of administration or viadiffering routes of administration.

PHARMACEUTICAL COMPOSITIONS

While it is possible for a compound of the present invention to beadministered alone, it is preferable to administer the compound as apharmaceutical formulation (composition). The compounds according to theinvention may be formulated for administration in any convenient way foruse in human or veterinary medicine. In certain embodiments, thecompound included in the pharmaceutical preparation may be activeitself, or may be a prodrug, e.g., capable of being converted to anactive compound in a physiological setting.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically acceptable dosage forms such as described below orby other conventional methods known to those of skill in the art.

Thus, another aspect of the present invention provides pharmaceuticallyacceptable compositions comprising a therapeutically effective amount ofone or more of the compounds described above, formulated together withone or more pharmaceutically acceptable carriers (additives) and/ordiluents. As described in detail below, the pharmaceutical compositionsof the present invention may be specially formulated for administrationin solid or liquid form, including those adapted for the following: (1)oral administration, for example, drenches (aqueous or non-aqueoussolutions or suspensions); tablets; boluses; powders; granules; pastesfor application to the tongue, teeth, lips, gums; mouth washes; gels;(2) parenteral administration, for example, by subcutaneous,intramuscular or intravenous injection as, for example, a sterilesolution or suspension; (3) topical application, for example, as acream, ointment or spray applied to the skin; (4) intravaginally orintrarectally, for example, as a pessary, cream or foam; or (5) forinhalation. However, in certain embodiments the subject compounds may besimply dissolved or suspended in sterile water. In certain embodiments,the pharmaceutical preparation is non-pyrogenic, i.e., does not elevatethe body temperature of a patient.

The phrase “therapeutically effective amount” as used herein means thatamount of a compound, material, or composition comprising a compound ofthe present invention which is effective for producing some desiredtherapeutic effect by inhibiting TRPV3 function in at least asub-population of cells in an animal and thereby blocking the biologicalconsequences of that function in the treated cells, at a reasonablebenefit/risk ratio applicable to any medical treatment.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” as used herein means apharmaceutically acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting the subject antagonistsfrom one organ, or portion of the body, to another organ, or portion ofthe body. Each carrier must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notinjurious to the patient. Some examples of materials which can serve aspharmaceutically acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalformulations.

As set out above, certain embodiments of the present compounds maycontain a basic functional group, such as amino or alkylamino, and are,thus, capable of forming pharmaceutically acceptable salts withpharmaceutically acceptable acids. The term “pharmaceutically acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ during the final isolation andpurification of the compounds of the invention, or by separatelyreacting a purified compound of the invention in its free base form witha suitable organic or inorganic acid, and isolating the salt thusformed. Representative salts include the hydrobromide, hydrochloride,sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate,palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate,glucoheptonate, lactobionate, and laurylsulphonate salts and the like.(See, for example, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm.Sci. 66:1-19)

The pharmaceutically acceptable salts of the subject compounds includethe conventional nontoxic salts or quaternary ammonium salts of thecompounds, e.g., from non-toxic organic or inorganic acids. For example,such conventional nontoxic salts include those derived from inorganicacids such as hydrochloride, hydrobromic, sulfuric, sulfamic,phosphoric, nitric, and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicyclic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isothionic, and the like.

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically acceptable salts with pharmaceutically acceptablebases. The term “pharmaceutically acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present invention. These salts can likewise beprepared in situ during the final isolation and purification of thecompounds, or by separately reacting the purified compound in its freeacid form with a suitable base, such as the hydroxide, carbonate orbicarbonate of a pharmaceutically acceptable metal cation, with ammonia,or with a pharmaceutically acceptable organic primary, secondary ortertiary amine. Representative alkali or alkaline earth salts includethe lithium, sodium, potassium, calcium, magnesium, and aluminum saltsand the like. Representative organic amines useful for the formation ofbase addition salts include ethylamine, diethylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine and the like. (See, forexample, Berge et al., supra)

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral,nasal, topical (including buccal and sublingual), rectal, vaginal and/orparenteral administration. Oral formulations include those delivered toand maintained in the mouth without swallowing, as well as formulationsthat are swallowed as part of or following use. The formulations mayconveniently be presented in unit dosage form and may be prepared by anymethods well known in the art of pharmacy. The amount of activeingredient which can be combined with a carrier material to produce asingle dosage form will vary depending upon the host being treated, theparticular mode of administration. The amount of active ingredient thatcan be combined with a carrier material to produce a single dosage formwill generally be that amount of the compound which produces atherapeutic effect. Generally, out of one hundred percent, this amountwill range from about 1 percent to about ninety-nine percent of activeingredient, preferably from about 5 percent to about 70 percent, mostpreferably from about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules and the like), theactive ingredient is mixed with one or more pharmaceutically acceptablecarriers, such as sodium citrate or dicalcium phosphate, and/or any ofthe following: (1) fillers or extenders, such as starches, lactose,sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as,for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol;(4) disintegrating agents, such as agar-agar, calcium carbonate, potatoor tapioca starch, alginic acid, certain silicates, and sodiumcarbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as, for example, cetyl alcohol and glycerolmonostearate; (8) absorbents, such as kaolin and bentonite clay; (9)lubricants, such a talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and(10) coloring agents. In the case of capsules, tablets and pills, thepharmaceutical compositions may also comprise buffering agents. Solidcompositions of a similar type may also be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugars, as well as high molecular weight polyethylene glycols andthe like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be sterilized by, for example,filtration through a bacteria-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved in sterile water, or some other sterile injectable mediumimmediately before use. These compositions may also optionally containopacifying agents and may be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain portion ofthe gastrointestinal tract, optionally, in a delayed manner Examples ofembedding compositions that can be used include polymeric substances andwaxes. The active ingredient can also be in micro-encapsulated form, ifappropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations for oral administration may be administered directly to themouth in the presence or absence of a device to aid administration orlocal delivery. For example, a liquid formulation or suspension may bedirectly delivered via a mouthwash. Alternatively, the liquidformulation or suspension may be directly applied to all or a portion ofthe mouth using a syringe or swab. In another embodiment, an oralformulation may be applied to a mouth guard or other dental device, anddelivered to the mouth via the mouth guard or device. The presentinvention contemplates that preparations suitable for oral delivery canbe formulated to facilitate any of these modes of delivery. For any ofthe foregoing, the oral formulation may optionally be ingested or may bemaintained in the mouth and later expectorated.

It is known that sterols, such as cholesterol, will form complexes withcyclodextrins. Thus, in preferred embodiments, where the inhibitor is asteroidal alkaloid, it may be formulated with cyclodextrins, such as α-,β- and γ-cyclodextrin, dimethyl-β cyclodextrin and2-hydroxypropyl-β-cyclodextrin.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such carriers as are known in theart to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants that may berequired.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the compoundin a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacteriostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents that delay absorption such as aluminum monostearate andgelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissue.

When the compounds of the present invention are administered aspharmaceuticals, to humans and animals, they can be given per se or as apharmaceutical composition containing, for example, 0.1 to 99.5% (morepreferably, 0.5 to 90%) of active ingredient in combination with apharmaceutically acceptable carrier.

The addition of the active compound of the invention to animal feed ispreferably accomplished by preparing an appropriate feed premixcontaining the active compound in an effective amount and incorporatingthe premix into the complete ration.

Alternatively, an intermediate concentrate or feed supplement containingthe active ingredient can be blended into the feed. The way in whichsuch feed premixes and complete rations can be prepared and administeredare described in reference books (such as “Applied Animal Nutrition”,W.H. Freedman and CO., San Francisco, U.S.A., 1969 or “Livestock Feedsand Feeding” O and B books, Corvallis, Ore., U.S.A., 1977).

Methods of introduction may also be provided by rechargeable orbiodegradable devices. Various slow release polymeric devices have beendeveloped and tested in vivo in recent years for the controlled deliveryof drugs, including proteinacious biopharmaceuticals. A variety ofbiocompatible polymers (including hydrogels), including bothbiodegradable and non-degradable polymers, can be used to form animplant for the sustained release of a compound at a particular targetsite.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compound employed, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound that is the lowest dose effective to producea therapeutic effect. Such an effective dose will generally depend uponthe factors described above. Generally, intravenous,intracerebroventricular and subcutaneous doses of the compounds of thisinvention for a patient will range from about 0.0001 to about 100 mg perkilogram of body weight per day.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms.

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

The compound of the invention can be administered as such or inadmixtures with pharmaceutically acceptable and/or sterile carriers andcan also be administered in conjunction with other agents. Exemplaryother agents include penicillins, cephalosporins, aminoglycosides,glycopeptides, anti-inflammatories, anti-virals, anti-fungals,anti-bacterials, or any agents appropriate for the treatment of theparticular injury, disease, or condition. Conjunctive therapy, thusincludes sequential, simultaneous and separate administration of theactive compound in a way that the therapeutic effects of the firstadministered compound are still detectable when the subsequent therapyis administered.

Synthetic Schemes and Identification of Active Antagonists CombinatorialLibraries

The compounds of the present invention, particularly libraries ofvariants having various representative classes of substituents, areamenable to combinatorial chemistry and other parallel synthesis schemes(see, for example, PCT WO 94/08051). The result is that large librariesof related compounds, e.g. a variegated library of compounds representedabove, can be screened rapidly in high throughput assays in order toidentify potential TRPV3 agonist or antagonist lead compounds, as wellas to refine the specificity, toxicity, and/or cytotoxic-kinetic profileof a lead compound. For instance, TRPV3 bioactivity assays, such asthose disclosed herein, can be used to screen a library of compounds forthose having agonist activity or antagonist activity towards TRPV3.

Simply for illustration, a combinatorial library for the purposes of thepresent invention is a mixture of chemically related compounds that maybe screened together for a desired property. The preparation of manyrelated compounds in a single reaction greatly reduces and simplifiesthe number of screening processes that need to be carried out. Screeningfor the appropriate physical properties can be done by conventionalmethods.

Diversity in the library can be created at a variety of differentlevels. For instance, the substrate aryl groups used in thecombinatorial reactions can be diverse in terms of the core aryl moiety,e.g., a variegation in terms of the ring structure, and/or can be variedwith respect to the other substituents.

A variety of techniques are available in the art for generatingcombinatorial libraries of small organic molecules such as the subjectcompounds. See, for example, Blondelle et al. (1995) Trends Anal. Chem.14:83; the Affymax U.S. Pat. Nos. 5,359,115 and 5,362,899: the EllmanU.S. Pat. No. 5,288,514: the Still et al. PCT publication WO 94/08051;the ArQule U.S. Pat. Nos. 5,736,412 and 5,712,171; Chen et al. (1994)JACS 116:2661: Kerr et al. (1993) JACS 115:252; PCT publicationsWO92/10092, WO93/09668 and WO91/07087; and the Lerner et al. PCTpublication WO93/20242). Accordingly, a variety of libraries on theorder of about 100 to 1,000,000 or more diversomers of the subjectcompounds can be synthesized and screened for particular activity orproperty.

Many variations on the above and related pathways permit the synthesisof widely diverse libraries of compounds that may be tested asinhibitors or agonists of TRPV3.

EXAMPLES Example 1 High-Throughput Screening Assay

The assay depends on detection of the rise in intracellular Ca²⁺concentration ([Ca²⁺]_(i)) following channel activation in cellsinducibly expressing the TRPV3 channel Ca²⁺ rise is quantified with theuse of fluorescent Ca²⁺ indicators that are loaded into cells andthereafter indicated the [Ca²⁺]_(i). Ca²⁺ influx follows activation ofthe TRPV3 channel Compounds inhibiting this [Ca²⁺]_(i) rise areconsidered hits for further investigation.

The commercially available HEK293/TREx line (Invitrogen) was stablytransfected with a TRPV3 construct and screened by immunostaining tofind clones with TRPV3 expression following stimulation with 1 μg/mltetracycline. Clonal TRPV3-expressing cells were maintained in thegrowth medium recommended by the manufacturer supplemented with 100μg/ml hygromycin to promote retention of the TRPV3 construct. Aftergrowing to near confluency, cells are plated at a density of ˜25,000cells/well in 384 well plates in the presence of 1 μg/ml tetracycline,and allowed to grow for 20-30 hrs. A nearly confluent monolayer results.Cells are then loaded with Ca²⁺ dye: Fura-2/AM or Fluo4/AM are added tothe wells to a final concentration of 2 μM or 1 respectively, andincubated for 80 mM or 60 mM, respectively, at room temperature.Supernatant is then removed from the cells by inverting plates with asharp flick, and 40 μl Ringer's solution (140 mM NaCl, 4.5 mM KCl, 2 mMCaCl₂, 1 mM MgCl₂, 10 mM HEPES, 10 mM glucose, pH 7.4) is then added toeach well. Following ˜1 hour for recovery from loading, cells areassayed using the Hamamatsu FDSS 6000 system, which permits illuminationalternately at 340 nM and 380 nM for Fura-2 experiments, or at 485 nMfor Fluo4 experiments. Frames were acquired at a rate of 0.2 Hz. Duringthe assay, the plates are continuously vortexed, with pipette mixing ofwells following addition of each reagent. For the screening assay, 13 μlof a diluted stock of each compound to be tested (at 50 μM) was added toeach well for 2 minutes following the collection of a short (4 frame)baseline. 13 μl 750 μM 2-APB (2-aminoethyldiphenylborinate) was added toeach well, achieving a final concentration of 10 μM each compound and150 μM 2-APB. Data were collected for ˜3 minutes following addition of2-APB, where the fluorescent intensity (for Fluo4) and the F340/F380ratio (for Fura-2) are proportional to the [Ca²⁺]_(i). Negative controlsconsisted of HEK293/TREx TRPV3 cells exposed to 2-APB, but no testcompound. Positive control cells were usually HEK293/TREx (“parental”)cells exposed to 2-APB but no test compound, but sometimes normalHEK/293 TREx TRPV3 cells were also used, but not exposed to 2-APB ortest compound. These controls defined a screening window, and “hits”were defined as those test compounds inhibiting the fluorescenceresponse by at least 40%.

Example 2 Patch Clamp Experiments

Whole-cell patch clamp experiments permit the detection of currentsthrough the TRPV3 channel in the cell line described above. A glasselectrode is brought into contact with a single cell and the membrane isthen ruptured, permitting control of the voltage of the cell membraneand measurement of currents flowing across the membrane using theamplifier attached to the electrode. A perfusion system permits controlof the extracellular solution, including the addition of blockers andactivators of the current. The current can be activated by heating thissolution to 28° C. or warmer or by addition of 20 μM 2-APB to thesolution.

TRPV3 cells were induced 20-48 hours, removed from growth plates, andreplated at low density (to attain good single-cell physical separation)on glass coverslips for measurement. In some cases, cells were grown inlow density overnight on glass coverslips. Patch clamp recordings weremade in the whole-cell mode with a holding potential of −40 mV. Every 5seconds, a voltage ramp was applied from −120 to +100 mV, 400 ms induration. Currents elicited were quantified at −80 mV and +80 mV. Theinternal solution consisted of 140 mM cesium aspartate, 10 mM EGTA, 2.27mM MgCl₂, 1.91 mM CaCl₂ and 10 mM HEPES, pH to 7.2 with KOH; with 50 nMcalculated free Ca^(2±). External solution was Ringer's solutiondescribed above. Upon addition of 2-APB or upon heating of theextraceullar solution as described above, TRPV3 current was induced onlyin TRPV3-expressing cells and not in parental HEK293 TREx cells. Thiscurrent showed a small inward component, reversal near +10 mV and astrong outward rectification, and is referred to as Phase I. Uponcontinued or repeated readdition of 2-APB or heat as a stimulus, currentcharacteristics change, resulting in a Phase II that is linear through+10 mV. Removal of the stimulus caused most of the current to go away,and inhibitor addition could still inhibit this current.

To determine whether compounds were selective for TRPV3 inhibition overinhibition of other ion channel types, the human ERG (hERG), NaV 1.2,and TRPV1 (hTRPV1) channels and the rat TRPV6 (rTRPV6) channel were alsostably transfected and expressed or induced to express in mammalian celllines. The methods for measuring currents from these channels arewell-established and have been described in numerous publications (See,Weerapura et al., 2002, J Physiology 540: 15-27; Rush et al., 2005, JPhysiology 564: 808-815; Caterina et al., 1997, Nature 389: 816-824;Hoenderhop et al., 2001, J Physiology 537: 747-761; Clapham et al.,2003, Pharmacol Rev 55: 591-596). Compounds of interest were testedagainst these channels at concentrations up to 30 μM, and the resultingdata were used to estimate IC₅₀.

Table 1 summarizes data collected for various tested compounds. The dataincludes approximate IC₅₀ values for inhibition of TRPV3 mediated inwardcurrent as assessed by patch-clamp.

TABLE 1 TRPV3 TRPV3 NaV1.2 Phase 1 Phase 2 TRPV1 Qpatch IC50 IC50 inwardinward hERG Cmpd inward inward IC50 IC50 IC50 ID Molecular Structure(nM) (nM) (nM) (nM) (nM)

≦5,000 ≦1,000 ≧30,000 6

≦500 ≦200 ≧20,000 ≧30,000 ≧30,000 21

≦5,000 ≦5,000 16

≦500 ≦500 ≧30,000 ≧5,000 20

≦200 ≦1,000 ≧30,000 ≧10,000 12

≦5,000 ≦10,000 8

≦1,000 ≦5,000 24

≦5,000 ≦10,000 10

≦200 ≦500

≦10,000 ≦5,000

≦200 ≦200

≦500 ≦500

≦5,000 ≦5,000

≦200 ≦200

≦10,000 ≦10,000

≦1,000 ≦500

≦500 ≦500

≦5,000 ≦1,000

≦200 ≦200

≦5,000 ≦5,000

Example 3 Other Screening Assays

Although the exemplary TRPV3 inhibitors provided herein were identifiedusing the assays described in Examples 1 and 2, other cell-based assayscan be used to identify and/or characterize TRPV3 inhibitors. One suchassay is described in U.S. application Ser. No. 11/078,188, filed Mar.11, 2005, the contents of which are hereby incorporated by reference intheir entirety. TRPV3 protein can be expressed in the prokaryotic cellsystem described in application Ser. No. 11/078,188, and this system canbe used to screen for compounds that modulate an activity of the TRPV3protein. Alternatively, an ion channel other than TRPV3 can be expressedin the prokaryotic cell system, and the system can be used to evaluatethe activity profile of an identified TRPV3 inhibitors with respect toother ion channels.

Any assays performed to identify and/or characterize compounds thatinhibit an activity of TRPV3 can be performed in a high-throughputfashion, or can be performed on a smaller scale examining individualcompounds or small numbers of compounds. Additionally, any of theseassays can be performed (i) as a primary assay to identify compoundsthat inhibit a function of TRPV3; (ii) as a secondary assay to assessthe specificity of a compound with respect to its activity against otherion channels; (iii) as an assay used in a medicinal chemistry program tooptimize subject compounds.

Example 4 Synthesis of Compounds of the Invention Synthesis of2-Chloro-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)ethanone (2)

2-Chloro-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)ethanone (2)

To a solution of 8-methyl-1,2,3,4-tetrahydroquinolin (1) (8.0 g, 54.3mmol) in dry CH₂Cl₂ (250 mL) at 0° C. was added chloroacetyl chloride(4.8 mL, 60.3 mmol) followed by addition of triethylamine (8.4 mL, 60.3mmol). The reaction mixture was stirred overnight under N₂ and thendiluted with CH₂Cl₂ (300 mL), washed with 1M HCl (2×225 mL), H₂O (2×250mL) and brine. Dried over Na₂SO₄, filtered and concentrated under reducepressure to 12.67 g of crude material which was purified by columnchromatography (EtOAc/Hexanes). Obtained 11.76 g (52.6 mmol) of target 2(off-white solid) in 97% yield.

Synthesis of 2-Chloro-1-(3,4-dihydro-2H-quinolin-1-yl)ethanone (4)

2-Chloro-1-(3,4-dihydro-2H-quinolin-1-yl)ethanone (4)

To a solution of 1,2,3,4-tetrahydroquinolin (3) (2.00 g, 15.0 mmol) indry CH₂Cl₂ (30 mL) at 0° C. was added chloroacetyl chloride (1.27 mL,16.0 mmol) followed by drop wise addition of triethylamine (2.50 mL,18.03 mmol). The reaction was stirred overnight under N₂. The reactionmixture was then diluted with CH₂Cl₂ (50 mL), washed with 5% HCl (15mL), H₂O (2×15 mL) and brine. Dried over MgSO₄, filtered andconcentrated to give 3.05 g of 4. This crude material was used insubsequent reactions without further purification.

Synthesis of2-(pyridine-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanones(6, 8, 10 and 12)

Synthesis of2-(5-trifluoromethyl-pyridine-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(6)

To a solution of 5-fluoromethyl-2-mercaptopyridine (5, 257 mg, 1.43mmol) in dry THF (9 mL) at 0° C. was added NaH (60% in mineral oil, 74.5mg, 1.9 mmol) in one portion. The reaction mixture was warmed to ambienttemperature and stirred for 30 min and then cooled back to 0° C.Compound 2 was added drop wise as a solution in THF (3 mL). The reactionwas stirred overnight at room temperature under N₂. The reaction wasquenched by addition of water (15 mL) and the product was extracted intoEtOAc (2×30 mL). The organic layer was washed with 20 mL of water,brine, dried over Na₂SO₄, filtered and concentrated to give 525 mg ofcrude material which was purified by Combi Flash chromatography(CH₂Cl₂/EtOAc). Obtained 300 mg (1.6 mmol) of target 6, as a pale yellowsolid, in 57% yield. MS (APCI) m/z: 367.1 (100%, [M+H]⁺, 220.0 (50%,[M-147+H]⁺; HPLC purity: 100%; Anal. Calcd. for C₁₈H₁₇F₃N₂OS: C, 59.00;H, 4.68; N, 7.65; F, 15.56; S, 8.75. Found: C, 58.73; H, 4.78; N, 7.72;F, 15.58; S, 8.83.

Synthesis of1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-2-(3-trifluoromethyl-pyridin-2-ylsulfanyl)-ethanone(8)

Compound 8 was synthesized according to procedure used for similarderivative 6. Starting with 3-fluoromethyl-2-mercaptopyridine (7, 200mg, 1.11 mmol) the target 8 was obtained as a yellow solid (150 mg, 38%yield). MS (APCI) m/z: 367.1 (100%, [M+H]⁺, 220.0 (20%, [M-147+H]⁺; HPLCpurity: 100%; Anal. Calcd. for C₁₈H₁₇F₃N₂OS: C, 59.00; H, 4.68; N, 7.65;F, 15.56; S, 8.75. Found: C, 59.13; H, 4.57; N, 7.50; F, 15.75; S, 8.82.

Synthesis of2-(3-chloro-5-trifluoromethyl-pyridine-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(10)

Compound 10 was synthesized according to procedure used for similarderivative 6. Starting with 3-chloro-5-fluoromethyl-2-mercaptopyridine(9, 300 mg, 1.4 mmol) the target 10 was obtained as a yellow oil (280mg, 50% yield). MS (APCI) m/z: 401.1 (100%, [M+H]⁺, 254.0 (15%,[M-147+H]⁺; HPLC purity: 100%; Anal. Calcd. for C₁₈H₁₆ClF₃N₂OS: C,53.93; H, 4.02; N, 6.99; Cl, 8.84; F, 14.22; S, 8.00. Found: C, 54.07;H, 3.98; N, 7.06; Cl. 8.89; F, 14.06; S, 8.04.

Synthesis of1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-2-(pyridine-2-ylsulfanyl)-ethanone(12)

Compound 12 was synthesized according to procedure used for similarderivative 6. Starting with 2-mercaptopyridine (11, 133 mg, 1.2 mmol)the target 12 was obtained as a pale yellow oil (130 mg, 33% yield). MS(APCI) m/z: 299.2 (100%, [M+H]⁺, 152.1 (20%, [M-147+H]⁺; HPLC purity:100%; Anal. Calcd. for C₁₇H₁₈N₂OS: C, 68.42; H, 6.08; N, 9.39; S, 10.75.Found: C, 68.20; H, 6.09; N, 9.20; S, 10.46.

Synthesis of2-(Chloro-pyridin-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanones(16, 20 and 21)

Synthesis of2-(5-Chloro-pyridin-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(16) 5-Chloro-pyridin-2-ylsulfanyl-acetic acid methyl ester (14)

To a solution of 2,5-dichloropyridine (13, 2.04 g, 13.8 mmol) in 40 mLof dry DMF was added K₂CO₃ (1.91 g, 13.8 mmol) and reaction mixture wasstirred for 30 min at ambient temperature. Neat methylthioglycolate(1.26 mL, 14 mmol) was added via syringe and the reaction mixture wasstirred overnight under N₂. The reaction mixture was treated with H₂O(120 mL) and the product was extracted into EtOAc (2×220 mL). Combinedorganic layers were washed with lot of water, brine, dried over Na₂SO₄,filtered and concentrated to give 2.3 g of crude material which waspurified by column chromatography (hexanes/EtOAc). Obtained 1.1 g (5.0mmol) of target 14, as a colorless oil, in 36% yield.

5-Chloro-pyridin-2-ylsulfanyl-acetic acid (15)

To the solution of the ester 14 (0.43 g, 2.0 mmol) in THF (18 mL) wasadded LiOH.H₂O (0.17 g, 4 mmol) as a solution in 5 mL of H₂O. Thereaction mixture was stirred at room temperature and monitored by TLC.When no staring material was observed, the reaction mixture wasacidified with 1N HCl, (˜4.5 mL) and concentrated under reduce pressureto remove the organic solvent. The mixture was diluted with 15 mL of H₂Oand extracted with EtOAc (2×120 mL). The organic layer was washed withbrine, dried over Na₂SO₄, concentrated and dried under high vacuum overP₂O₅ to give 0.40 g (98% yield) of compound 15 which was used insubsequent reaction without further purification.

2-(5-Chloro-pyridin-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(16)

To a solution of (5-chloro-pyridin-2-ylsulfanyl)-acetic acid (15, 0.27g, 1.3 mmol) in dry CH₂Cl₂ (10 mL) and DMF (2 mL) at 0° C. was addedHATU (0.5 g, 1.3 mmol) and reaction mixture was stirred for 30 min atambient temperature. Then the reaction mixture was cooled to 0° C. andneat 8-methyl-1,2,3,4-tetrahydroquinolin (0.20 g, 1.3 mmol) was addedvia syringe followed by addition of DIPEA, (0.69 mL, 3.9 mmol). Thereaction mixture was allowed to warm to room temperature over 18 h. Thereaction mixture was diluted with 50 mL of CH₂Cl₂, washed with H₂O, 10%citric acid, NaHCO₃ (saturated), brine, dried over Na₂SO₄ andconcentrated. The crude was purified by column chromatography(hexanes/EtOAc) to give 90 mg of the target 16 as a white solid in 20%yield. MS (APCI) m/z: 333.1 (100%, [M+H]⁺, 186.0 (20%, [M-147+H]⁺; HPLCpurity: 100%; Anal. Calcd. for C₁₇H₁₇ClN₂OS: C, 61.34; H, 5.15; N, 8.42;Cl, 10.65; S, 9.63. Found: C, 61.33; H, 5.06; N, 8.36; Cl. 10.67; S,9.56.

Synthesis of2-(3-chloro-pyridin-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(20)

(3-Chloro-pyridin-2-ylsulfanyl)-acetic acid methyl ester (18) wassynthesized according to the procedure used for compound 14. Startingwith 2,3-dichloropyridine (17, 2.09 g, 14.0 mmol) the target 18 wasobtained as a colorless oil (1.6 g, 52% yield).

(3-Chloro-pyridin-2-ylsulfanyl)-acetic acid (19) was synthesizedaccording to the procedure used for compound 15. Starting with 0.69 g(3.2 mmol) of corresponding ester 18 the target 19 was obtained as awhite solid (0.63 g) in 97% yield.

2-(3-Chloro-pyridin-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(20)

To a solution of (3-chloro-pyridin-2-ylsulfanyl)-acetic acid (19, 0.27g, 1.3 mmol) in 12 mL of dry THF at 0° C. was added neat8-methyl-1,2,3,4-tetrahydroquinolin (0.20 g, 1.3 mmol) followed byaddition of DMAP (0.19 g, 1.6 mmol) and EDCI (0.31 g, 1.6 mmol). Thereaction was stirred for 20 h at room temperature under N₂. The reactionmixture was diluted with 50 mL of CH₂Cl₂, washed with H₂O, 10% citricacid, NaHCO₃ (saturated), brine, dried over Na₂SO₄ and concentrated. Thecrude was purified by column chromatography (hexanes/EtOAc) to give 0.15g (33% yield) of the target 20 as an off-white solid. MS (APCI) m/z:333.1 (100%, [M+H]⁺, 186.0 (35%, [M-147+H]⁺; HPLC purity: 100%; Anal.Calcd. for C₁₇H₁₇ClN₂OS: C, 61.34; H, 5.15; N, 8.42; Cl, 10.65; S, 9.63.Found: C, 61.04; H, 5.28; N, 8.36; Cl. 10.78; S, 9.68.

Synthesis of2-(5-chloro-pyridine-2-ylthio)-1-(3,4-dihydroquinolin-1-(2H)-yl)-ethanone(21)

The compound 21 was obtained according to the procedure used for thederivative 20 but using base (DIPEA) as well:(3-chloro-pyridin-2-ylsulfanyl)-acetic acid (15, 387 mg, 1.9 mmol),DIPEA (0.66 mL, 3.8 mmol), 1,2,3,4-tetrahydroquinolin (3) (0.24 mL g,1.9 mmol), DMAP (0.27 g, 2.2 mmol) EDCI (0.42 g, 2.2 mmol). The target21 was obtained as a white solid (0.32 mg, 53% yield). MS (APCI) m/z:319.1 (70%, [M+H]⁺, 186.0 (35%, [M-133+H]⁺; HPLC purity: 100%; Anal.Calcd. for C₁₆H₁₅ClN₂OS: C, 60.28; H, 4.74; N, 8.79; Cl, 11.12; S,10.06. Found: C, 59.99; H, 4.75; N, 8.70; Cl. 11.36; S, 10.03.

Synthesis of1-(8-Methyl-3,4-dihydro-2H-quinolin-1-yl)-2-(5-methyl-pyridin-2-ylsulfanyl)-ethanone(24)

5-Methyl-pyridine-2-thiol (23)

2-bromo-5-methyl-pyridine (22, 1.61 g, 9.4 mmol) was reacted withNaHS.H₂O (5.5 g, 75.8 mmol) in 15 mL of 1,2-propylenediol in a sealedtube at 160° C. After 1.5 h no starting material was detected by TLC.Cooled to room temperature and 1.2 mL of glacial acetic acid was addeddrop wise to the reaction mixture and the 5-methyl-pyridine-2-thiol (23)formed as a pale yellow powder. The crude was purified by Combi Flashchromatography (CHCl₃/CH₃CN). Obtained 0.25 g (2.0 mmol) of target 23,as a pale yellow solid, in 21% yield.

1-(8-Methyl-3,4-dihydro-2H-quinolin-1-yl)-2-(5-methyl-pyridin-2-ylsulfanyl)-ethanone(24) was obtained according to the procedure used for a similar compound(6). Starting with 120 mg (0.96 mmol) of corresponding thiol (23,5-methyl-pyridine-2-thiol) 150 mg (50% yield) of the target 24 wasobtained as a white solid. MS (APCI) m/z: 313.2 (100%, [M+H]⁺, 166.1(35%, [M-147+H]⁺; HPLC purity: 99.2%; Anal. Calcd. for C₁₈H₂₀N₂OS: C,69.20; H, 6.45; N, 8.97; S, 10.26. Found: C, 69.19; H, 6.53; N, 8.89; S,10.31.

Example 5 AMPA Receptor Binding Study

In certain embodiments, compounds of the invention inhibit aTRPV3-mediated current with a particular IC₅₀ and, at a concentrationequal to that IC₅₀, the compounds do not appreciably bind the AMPAreceptor (e.g., do not specifically bind the AMPA receptor and/ordisplace a more than 10% of a specifically bound, high affinity ligand).In certain embodiments, compounds of the invention inhibit aTRPV3-mediated current with an IC₅₀ that is more potent than its Ki forthe AMPA receptor. An exemplary assay for evaluating binding ofcompounds to the AMPA receptor is summarized in the following: Murphy etal., 1987, Neurochem Res 12: 775-781 and Morgan et al., 1991, NeurochemInt 18: 75-84.

Briefly, binding curves are generated by quantifying the displacement ofa radiolabelled AMPA ligand in the presence of a compound of interest.For example, binding curves are generated using high affinity compoundssuch as AMPA ligand or quisqualic acid. Binding curves using lowaffinity reference compounds such as NMDA or kainic acid may also begenerated. From these binding curves a binding constant and/or bindingaffinity for a reference can be ascertained. From these binding curves,and based on whether a compound of interest (e.g., a TRPV3 inhibitorcompound) displaces appreciable amounts of a high affinity AMPA receptorbinding compound, a binding constant and/or binding affinity for a TRPV3inhibitor compound can be ascertained.

Binding curves for TRPV3 antagonists can be similarly generated toascertain whether a TRPV3 antagonist of the present inventionappreciably binds the AMPA receptor. For a given TRPV3 antagonist forwhich the IC₅₀ for inhibiting a TRPV3-mediated current is known, one canreadily ascertain whether the compound appreciably binds an AMPAreceptor at a concentration equivalent to its IC₅₀ for inhibiting aTRPV3-mediated current. Furthermore, one can ascertain a Ki anddetermine whether the Ki for binding the AMPA receptor is less potentthan the IC₅₀ for inhibiting a TRPV3-mediated current. Furthermore, abinding constant and/or affinity concentration of a compound for theAMPA receptor can be determined.

Example 6 Testing of TRPV3 Antagonists in a Formalin Model of Pain

As outlined above, the formalin model involves injection of a formalinsolution intradermally or intraperitoneally. Injection of formalinsolution invokes a biphasic response, and thus provides a model for bothnociceptive and inflammatory pain. The formalin model can be used toevaluate the effectiveness of an exemplary TRPV3 inhibitor in thetreatment of pain.

Briefly, the following protocol can be followed. Male Holtzmann rats aregiven intraplantar injections of 50 μL of 2% formalin. Paw flinching isdetected by an automated sensor detecting movement of a small metal bandplaced on the injected hind paw. Drug or vehicle is administeredapproximately 15 minutes prior to the injection of formalin. Theanimal's response to injection of the irritant is measured by countingflinches per minutes during the Early Phase (the first 5 minutesfollowing injection of formalin), during the Late Phase (approximately30 minutes after injection of formalin), and during the intervening painfree phase.

Note that efficacy of various TRPV3 inhibitors can be evaluatedfollowing administration via any of a number of routes (oral, IP, IV,etc) and at any of a number of doses. Efficacy can be compared tovehicle control drug and/or efficacy can be compared to knownpain-reducing medicaments.

Example 7 Testing of TRPV3 Antagonists in the CFA Model of InflammatoryPain

As outlined above, the Complete Freund's Adjuvant (CFA) model is a modelof inflammatory pain. As such, it may be used to evaluate effectivenessin relieving pain caused by inflammation, for example, pain due toarthritis and other inflammatory conditions.

Naive rats are pretested for sensitivity to a cold or mechanicalstimulus. The next day, 100 μL of complete Freund's adjuvant (CFA) isinjected into the plantar surface of the right hindpaw. Two days later,in the morning, the rats are again pretested. In the afternoon, rats areinjected with either vehicle control or with drug. Drugs or vehicle areinjected intraperitoneally, and 45 minutes later rats are tested forhyperalgesia by applying the cold source or Von Frey filament to the CFAinjected and uninjected hindpaw and measuring latency to withdrawal.

Note that efficacy of various TRPV3 inhibitors can be evaluatedfollowing administration via any of a number of routes (oral, IP, IV,etc) and at any of a number of doses. Efficacy can be compared tovehicle control drug and/or efficacy can be compared to knownpain-reducing medicaments.

Example 8 Testing of TRPV3 Antagonists in a Thermal Injury Model of Pain

The thermal injury model can be used to evaluate the effectiveness of anexemplary TRPV3 inhibitor in the treatment of nociceptive pain.

Briefly, the following protocol may be used. Male Holtzman rats(approximately 300 grams) are tested on thermal escape using aHargreaves type apparatus. Under light anesthesia, a thermal injury (52°C. for 45 seconds) is applied to one heel. The animals are tested forthermal escape latency of the injured and uninjured paw before and at30, 60, 80, and 120 minutes after injury. Drug (a TRPV3 inhibitor) orvehicle (0.5% methylcellulose) is administered after the baselinemeasurement and approximately 15-20 minutes prior to the thermal injury.In addition to the escape latency measurement, behavioral observationsare made throughout the experiment.

Note that efficacy of various TRPV3 inhibitors can be evaluatedfollowing administration via any of a number of routes (oral, IP, IV,etc) and at any of a number of doses. Efficacy can be compared tovehicle control drug and/or efficacy can be compared to knownpain-reducing medicaments.

Example 9 Testing of TRPV3 Antagonists in the Chung Model of NeuropathicPain

Briefly, male Sprague Dawley rats (approximately 175 grams) are preparedwith ligation of the L4/5 nerve roots. After 5-8 days, the animals aretested for tactile allodynia using Von Frey hairs. Thresholds areassessed with the “up-down” method. Drug or vehicle is administered andthe animals tested periodically over the next four hours.

Note that efficacy of various TRPV3 inhibitors can be evaluatedfollowing administration via any of a number of routes (oral, IP, IV,etc) and at any of a number of doses. Efficacy can be compared tovehicle control drug and/or efficacy can be compared to knownpain-reducing medicaments.

Example 10 Testing of TRPV3 Antagonists in the Carrageenan Model ofAcute Inflammatory Pain

As outlined above, the carrageenan model is a model of acuteinflammatory pain. As such, it may be used to evaluate effectiveness inrelieving pain caused by inflammation, for example, pain due toarthritis.

Briefly, naive rats are pretested for sensitivity to a heat stimulususing the Hargreaves apparatus. The next day, 100 μL of 2-carrageenan isinjected into the plantar surface of the right hindpaw approximately 4.5hours before testing. 30-60 minutes before testing the rats are injectedadministered vehicle or drug. Following administration of bothcarrageenan and drug or vehicle control, the thermal escape latency ismeasured. Data can be expressed as a percentage comparing the recordedPaw Withdrawal Latencies (PWLs) in seconds to that pre-carrageenanadministration.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein, are hereby incorporatedby reference in their entirety as if each individual publication orpatent was specifically and individually indicated to be incorporated byreference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

1-38. (canceled)
 39. A method for treating allergies in a subject, themethod comprising administering to the subject a compound of formula Ior a pharmaceutically acceptable salt, solvate, hydrate, oxidativemetabolite, or prodrug thereof:

wherein: R₁, R₂, R₃, and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; mrepresents 0, 1, 2, 3, or 4; and R₅ represents a substituent on the ringto which it is attached.
 40. The method of claim 39, wherein each of R₁,R₂, R₃ and R₄ represent H, lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, nitro, halogen, CF₃, aryl, —CH₂CH═CF₂,—CH═CF₂, or cyano.
 41. The method of claim 39, wherein R₅ is,independently for each occurrence, selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, andcyano.
 42. The method of claim 39, wherein X is S; R₁ represents H; R₂,R₃ and R₄ represent H, lower alkyl, halogen, or CF₃; and m is
 0. 43. Themethod of claim 39, wherein the compound of formula I is a compound offormula Ib or a pharmaceutically acceptable salt, solvate, hydrate,oxidative metabolite, or prodrug thereof

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; yrepresents 0, 1, 2, or 3; R₅ represents a substituent on the ring towhich it is attached; R₅′ represents a substituent.
 44. The method ofclaim 39, wherein the compound of formula I is


45. A method for treating a gastrointestinal disease in a subject, themethod comprising administering to the subject a compound of formula Ior a pharmaceutically acceptable salt, solvate, hydrate, oxidativemetabolite, or prodrug thereof:

wherein: R₁, R₂, R₃, and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; mrepresents 0, 1, 2, 3, or 4; and R₅ represents a substituent on the ringto which it is attached.
 46. The method of claim 45, wherein thegastrointestinal disease Crohn's disease, ulcerative colitis, irritablebowel syndrome, celic sprue, or dermatitis herpetiformis.
 47. The methodof claim 45, wherein each of R₁, R₂, R₃ and R₄ represent H, lower alkyl,alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl,hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro, halogen,CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano.
 48. The method of claim 45,wherein R₅ is, independently for each occurrence, selected from loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, and cyano.
 49. The method of claim 45, wherein X is S; R₁represents H; R₂, R₃ and R₄ represent H, lower alkyl, halogen, or CF₃;and m is
 0. 50. The method of claim 45, wherein the compound of formulaI is a compound of formula Ib or a pharmaceutically acceptable salt,solvate, hydrate, oxidative metabolite, or prodrug thereof:

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; yrepresents 0, 1, 2, or 3; R₅ represents a substituent on the ring towhich it is attached; R₅′ represents a substituent.
 51. The method ofclaim 45, wherein the compound of formula I is


52. A method for treating an inflammatory disease or disorder in asubject, the method comprising administering to the subject a compoundof formula I or a pharmaceutically acceptable salt, solvate, hydrate,oxidative metabolite, or prodrug thereof:

wherein: R₁, R₂, R₃, and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; mrepresents 0, 1, 2, 3, or 4; and R₅ represents a substituent on the ringto which it is attached.
 53. The method of claim 52, wherein theinflammatory disease or disorder is asthma, chronic obstructivepulmonary disease, rheumatoid arthritis, osteoarthritis, inflammatorybowel disease, glomerulonephritis, or multiple sclerosis.
 54. The methodof claim 52, wherein each of R₁, R₂, R₃ and R₄ represent H, lower alkyl,alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl,hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro, halogen,CF₃, aryl, —CH₂CH═CF₂, —CH═CF₂, or cyano.
 55. The method of claim 52,wherein R₅ is, independently for each occurrence, selected from loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, and cyano.
 56. The method of claim 52, wherein X is S; R₁represents H; R₂, R₃ and R₄ represent H, lower alkyl, halogen, or CF₃;and m is
 0. 57. The method of claim 52, wherein the compound of formulaI is a compound of formula Ib or a pharmaceutically acceptable salt,solvate, hydrate, oxidative metabolite, or prodrug thereof

wherein: R₁, R₂, R₃ and R₄ represent H or a substituent; X represents Sor O; n represents 1 or 2 when X is O, or n represents 1 when X is S; yrepresents 0, 1, 2, or 3; R₅ represents a substituent on the ring towhich it is attached; R₅′ represents a substituent.
 58. The method ofclaim 52, wherein the compound of formula I is